Treatment of GVHD

ABSTRACT

The invention relates to treatment of graft versus host disease (GVHD) using compounds that inhibit ROCK2. In preferred aspects, the present invention provides methods for the treatment of GVHD, including chronic GVHD (cGVHD) using compounds having the formulae 1-XXV, as set forth herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Application No. 61/977,564,filed on Apr. 9, 2014, which is incorporated herein by reference in itsentirety.

FIELD OF THE INVENTION

The invention relates to treatment of graft versus host disease (GVHD)using compounds that inhibit ROCK2.

BACKGROUND OF THE INVENTION

Rho-associated protein kinase (ROCK) is a key intracellular regulator ofcytoskeletal dynamics and cell motility. Rho-kinase regulates a numberof downstream targets of RhoA through phosphorylation, including, forexample, myosin light chain, the myosin light chain phosphatase bindingsubunit and LIM-kinase 2. These substrates regulate actin filamentorganization and contractility. In smooth muscle cells Rho-kinasemediates calcium sensitization and smooth muscle contraction. Inhibitionof Rho-kinase blocks 5-HT and phenylephrine agonist induced musclecontraction. When introduced into non-smooth muscle cells, Rho kinaseinduces stress fiber formation and is required for the cellulartransformation mediated by RhoA. Rho kinase participates in a variety ofcellular processes, including but not limited to cell adhesion, cellmotility and migration, growth control, cell contraction, andcytokinesis. Rho kinase is also involved in Na/H exchange transportsystem activation, stress fiber formation, adducin activation, andphysiological processes such as vasoconstriction, bronchial smoothmuscle constriction, vascular smooth muscle and endothelial cellproliferation, platelet aggregation, and others.

Inhibition of Rho-kinase activity in animal models has demonstrated anumber of benefits of Rho-kinase inhibition for the treatment of humandiseases. These include models of cardiovascular diseases such ashypertension, atherosclerosis, restenosis, cardiac hypertrophy, ocularhypertension, cerebral ischemia, cerebral vasospasm, penile erectiledysfunction, central nervous system disorders such as neuronaldegeneration and spinal cord injury, and in neoplasias. Inhibition ofRho-kinase activity has been shown to inhibit tumor cell growth andmetastasis, angiogenesis, arterial thrombotic disorders such as plateletaggregation and leukocyte aggregation, asthma, regulation ofintraoccular pressure, and bone resorption. The inhibition of Rho-kinaseactivity in patients has benefits for controlling cerebral vasospasmsand ischemia following subarachnoid hemorrhage, reduction of intraocularpressure, increase in ocular aqueous outflow by relaxation of trabecularmeshwork tissue, improving blood flow to the optic nerve, and protectionof healthy ganglion cells.

In mammals, Rho-kinase consists of two isoforms, ROCK1 (ROCKβ;p160-ROCK) and ROCK2 (ROCKα). ROCK1 and ROCK2 are differentiallyexpressed and regulated in specific tissues. For example, ROCK1 isubiquitously expressed at relatively high levels, whereas ROCK2 ispreferentially expressed in cardiac and brain and skeletal muscle. Theisoforms are also expressed in some tissues and in a developmental stagespecific manner. ROCK1 is a substrate for cleavage by caspase-3 duringapoptosis, whereas ROCK2 is not. Smooth muscle specific basic calponinis phosphorylated only by ROCK2.

Given the extent of involved cellular processes and diseases, compoundsthat selectively inhibit one rho kinase, or inhibit ROCK1 and ROCK2, aredesired.

SUMMARY OF THE INVENTION

The present invention relates to treatment of graft versus host disease(GVHD), including chronic GVHD (cGVHD) using compounds having theformulae I-XXV, as set forth herein. In certain embodiments, theinvention provides a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   -   X is selected from N or C—R₁;    -   Y is selected from N or C—R₅;    -   Z is selected from N or C—R₃;    -   R₁ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₂ is a group having the formula -A-R₁₀;    -   A is selected from the group consisting of a covalent bond,        aryl, heteroaryl, cycloalkyl, and heterocyclyl;    -   R₁₀ is selected from the group consisting of H, CN, halo,        hydroxy, lower alkoxy, amino, perfluoro lower alkyl, C₁-C₁₀        alkyl, C₂-C₁₀ alkenyl, and -(M)_(x)-(CH₂)_(y)—R₁₁;    -   M is selected from the group consisting of N—R₂₀, CR₂₁R₂₂, and        C═O;    -   x is 0 or 1;    -   R₂₀ is selected from H and C₁₋₅ alkyl;    -   R₂₁ and R₂₂ are independently selected from the group consisting        of H, halogen, and lower alkyl, or alternatively R₂₁ and R₂₂ may        be taken together with the atom to which they are attached to        form a C₃₋₆ cycloalkyl;    -   y is 0, 1, 2, 3, 4, 5, or 6;    -   R₁₁ is selected from the group consisting of H, C₁₋₆ alkyl,        optionally substituted C₃₋₆ cycloalkyl, optionally substituted        aryl, optionally substituted heteroaryl, optionally substituted        heterocyclyl, wherein the optional substituents are selected        from the group consisting of lower alkyl, C₁₋₆ cycloalkyl, oxo,        CN, halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl,        and (lower alkyl)-O-(lower alkyl);    -   alternatively R₁₁ is selected from the group consisting of        —NR₁₃R₁₄, —C(═O)NR₁₃R₁₄, and —C(═O)R₁₂, and —CO₂R₁₂;    -   R¹² is selected from the group consisting of C₁-C₁₀ alkyl, aryl,        heteroaryl, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), aralkyl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, oxo, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆ alkoxy,        (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), hydroxy, cyano and C₁-C₃        perfluoro alkyl;    -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₆        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, oxo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl,        C₁-C₆ alkoxy, (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), hydroxy, amino,        cyano and C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), oxo, hydroxy,        cyano and C₁-C₃ perfluoro alkyl;    -   R₃ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₅ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₆ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₇ is selected from the group consisting of H and lower alkyl;        and    -   R₈ is a nitrogen-containing heterocyclic ring system ring which        may comprise 0-2 additional ring heteroatoms selected from N, O        and S, and may be unsubstituted or may be substituted with 1 to        3 substituents selected from halo, CN, oxo, hydroxy, amino,        lower alkyl, perfluoro lower alkyl, and lower alkoxy.

In further embodiments, the present invention relates to treatment ofGVHD, such as cGVHD using a compound having the formulae XXVI:

or pharmaceutically acceptable salt thereof, wherein:R¹ is selected from the group consisting of H or C₁-C₆ alkyl;R² is selected from the group consisting of aryl, heteroaryl, C₃-C₇cycloalkyl, a three to twelve membered heterocyclic ring containing upto 3 heteroatoms, each of which may be optionally substituted from 1 to3 substituents independently selected from halo and C₁-C₆ alkyl;

-   -   X is selected from N or CR³;    -   Y is selected from N or CR³;    -   Z is selected from N or CR⁴;    -   W is selected from CR⁵;    -   wherein X, Y, and Z are independently selected and at least one        of which is N;        wherein each R³ is independently selected from the group        consisting of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl,        —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        R⁴ and R⁵ are independently selected from the group consisting        of H, halo, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₁-C₆        alkoxy, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR⁴¹R⁴²,        —(CH₂)_(x)NR⁴¹R⁴² and —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl;    -   R⁴¹ and R⁴² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   or R⁴¹ and R⁴² may be taken together to form a three to twelve        membered cycloalkyl or heterocyclic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo, C₁-C₆ alkyl, and        C₁-C₆ alkoxy;    -   x is selected from 0 to 6;        or R⁴ and R⁵ may be taken together to form a three to twelve        membered heterocyclic or aromatic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo and C₁-C₆ alkyl        and C₁-C₆ alkoxy;    -   a is selected from 0 to 2;    -   b is selected from 0 to 2;    -   wherein a or b are independently selected and one of which is at        least 1        A is a three to twelve membered heterocyclic or aromatic ring        having up to 3 heteroatoms which is optionally substituted from        1 to 3 substituents independently selected from halo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, C₃-C₇ cycloalkyl, oxo, acyl,        —S—(C₁-C₆ alkyl), OH, NH₂, CN and C₁-C₃ perfluroalkyl.

In other embodiments, the present invention relates to treatment ofGVHD, such as cGVHD using a compound having the formula XXXI:

or pharmaceutically acceptable salt, wherein:R¹ is selected from the group consisting of —O—(CH₂)_(y)—CO₂R¹²,—O—(CH₂)_(y)—C(═O)NR¹³R¹⁴, —O—(CH₂)_(y)-heteroaryl,—O—(CH₂)_(y)-cycloalkyl, —O—C(═O)—(CH₂)_(y)—NR¹³R¹⁴,—O—(CH₂)_(z)—NR¹³R¹⁴, —NH—C(═O)—(CH₂)_(y)—NR¹³R¹⁴, —NH—C(═O)—X—R¹⁵,—NH—(CH₂)_(y)—NR¹³R¹⁴;R¹² is selected from the group consisting of C₁-C₆ alkyl, —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆alkyl)-C(═O)NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted at one or more carbon atoms by from 1 to 3substituents independently selected from halo, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆        alkyl)-C(═O)NR¹⁶R¹⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, OH, NH₂, CN and C₁-C₃ perfluoro alkyl;    -   X is selected from a covalent bond, O, NH, and C₁-C₆ alkyl;    -   R¹⁵ is selected from the group consisting of heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, OH, NH₂, CN        and C₁-C₃ perfluoro alkyl;    -   or R¹⁵ is selected from —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆        alkyl)-NR¹⁶R¹⁷, —CO₂R¹⁸, —O—(CH₂)_(x)—CO₂R¹⁸, and —C(═O)NR¹⁶R¹⁷;        -   R¹⁶ and R¹⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, OH, NH₂, CN and C₁-C₃ perfluoro            alkyl;        -   or R¹⁶ and R¹⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R¹⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;    -   x is selected from 0 to 6;    -   y is selected from 0 to 6;    -   z is selected from 2 to 6;        each R² is independently selected from the group consisting of        lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and        perfluoro lower alkyl;        each R³ is independently selected from the group consisting of        lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and        perfluoro lower alkyl;        R⁴ is selected from —(CH₂)_(a)—NR⁴³R⁴⁴, —Y—R⁴²,        —O—(CH₂)_(a)—CO₂R⁴², —O—(CH₂)_(a)—C(═O)NR⁴³R⁴⁴,        —O—(CH₂)_(a)-heteroaryl, —O—(CH₂)_(a)-cycloalkyl,        —O—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴, —O—(CH₂)_(c)—NR⁴³R⁴⁴,        —NH—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴, —NH—C(═O)—Y—R⁴⁵,        —NH—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴;    -   R⁴² is selected from the group consisting of C₁-C₆ alkyl,        —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆        alkyl)-C(═O)NR⁴⁶R⁴⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆        alkyl), each of which may be optionally substituted at one or        more carbon atoms by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and        C₁-C₃ perfluoro alkyl;    -   R⁴³ and R⁴⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆        alkyl)-C(═O)NR⁴⁶R⁴⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁴³ and R⁴⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   Y is selected from a covalent bond, O, NH, and C₁-C₆ alkyl;    -   R⁴⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —CO₂R⁴⁸,        —O—(CH₂)_(b)—CO₂R⁴⁸, and —C(═O)NR⁴⁶R⁴⁷,        -   R⁴⁶ and R⁴⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁴⁶ and R⁴⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁴⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;    -   a is selected from 0 to 6;    -   b is selected from 0 to 6;    -   c is selected from 2 to 6;        R⁵ is selected from the group consisting of H, C₁-C₆ alkyl,        —(CH₂)_(d)—C(═O)—NR⁵³R⁵⁴, —C(═O)—(CH₂)_(d)—NR⁵³R⁵⁴,        —C(═O)—X—R⁵⁵, and —C(═O)—(CH₂)_(d)—NR⁵³R⁵⁴;    -   R⁵³ and R⁵⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —(C₁-C₆        alkyl)-C(═O)NR⁵⁶R⁵⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁵³ and R⁵⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro        alkyl;    -   R⁵⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —CO₂R⁵⁸,        —O—(CH₂)_(e)—CO₂R⁵⁸, and —C(═O)NR⁵⁶R⁵⁷,        -   R⁵⁶ and R⁵⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁵⁶ and R⁵⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁵⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;        -   d is selected from 0 to 6;        -   e is selected from 0 to 6;            R⁶ is selected from the group consisting of H, C₁-C₆ alkyl,            —(CH₂)_(r)—C(═O)—NR⁶³R⁶⁴, —C(═O)—(CH₂)_(r)—NR⁶³R⁶⁴,            —C(═O)—X—R⁶⁵, and —C(═O)—(CH₂)_(r)—NR⁶³R⁶⁴;    -   R⁶³ and R⁶⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —(C₁-C₆        alkyl)-C(═O)NR⁶⁶R⁶⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁶³ and R⁶⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro        alkyl;    -   R⁶⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —CO₂R⁶⁸,        —O—(CH₂)_(s)—CO₂R⁶⁸, and —C(═O)NR⁶⁶R⁶⁷,        -   R⁶⁶ and R⁶⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁶⁶ and R⁶⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁶⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;    -   r is selected from 0 to 6;    -   s is selected from 0 to 6;        n is selected from 0 to 4;        m is selected from 0 to 3; and        p is selected from 0 and 1.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows compounds of the invention.

FIG. 2 shows compounds of the invention.

FIG. 3 shows compounds of the invention.

FIG. 4 shows compounds of the invention.

FIG. 5 shows compounds of Formula XXVI of the invention.

FIG. 6 shows dose response curves for inhibition of ROCK1 vs. ROCK2.Compounds correspond to Examples herein, as follows: K100-5, Ex. 12;KD-25, SLx-2119; 3272, Ex. 28; K100-04, Ex. 14; K100-16, Ex. 43;K100-21; Ex. 38; K100-23 Ex. 52; K100-24, Ex. 111; K100-25, Ex. 56;K100-26, Ex. 13; 3266, Ex. 26.

FIG. 7 compares ROCK1 and ROCK2 inhibition among the compounds ofExamples 43, 48, and 118.

FIG. 8 shows ROCK2 selective inhibitor, KD025 (SLx 2119), inhibitsIL-17/IL-21 secretion (A) and proliferation (B) in human CD4⁺ T cells invitro.

FIG. 9 shows ROCK2 siRNA, but not ROCK1 siRNA, inhibits IL-17 and IL-21secretion. Panel A, left: Anti-ROCK1 siRNA reduced Rock1 expression byabout 75%. Anti-ROCK2 siRNA reduced Rock2 expression by about 85%. PanelA, right: ROCK2 siRNA, but not ROCK1 siRNA, inhibited IL-17 and IL-21expression. No inhibition of IFN-γ was observed. Panel B: ROCK2 siRNA,but not ROCK1 siRNA, inhibited phosphorylation of Stat3, IRF4, andRORγt. Panel C: ROCK2 siRNA, but not ROCK1 siRNA, inhibitedphosphorylation of MLC.

FIG. 10 shows compound 181 of the invention regulates cytokinesecretion. Panel A shows the compound of Example 181 inhibits secretionof IL-21, but not INF-γ or IL-2. Panel B shows inhibition of IL-17secretion is dose-dependent. Legend (A and B): left bars 1: KD025 (aROCK2 selective inhibitor); center bar: compound of Example 181dissolved in HCL; right bar; compound of Example 181 dissolved in DMSO.

FIG. 11 shows KD025 (SLx 2119) inhibits STAT3 phosphorylation. (A)Pre-treatment of T cells with KD025 followed by stimulation withanti-CD3/CD28 antibodies. (B) Cell culture under Th17-skewing conditionsfor 5 days followed by treatment with KD025 for 3 hours. (C) CD4⁺ Tcells were activated by anti-CD3/CD28, TGF-β and IL-1β with 0 μM, 2.5μM, 5 μM, or 10 μM KD025 for 48 hours

FIG. 12 shows ROCK2 selective inhibitor, KD025, inhibits IL-17, IL-21and IFN-γ production ex vivo in CD3/CD28 stimulated CD4⁺ T cells from RApatients. Panel A: In RA patients, KD025 inhibits TCR stimulation ofIL-17, and IL-21, as well as IFN-γ. Panel B: Inhibition of IFN-γproduction is correlated with disease activity score (DAS). Panel C:Frequency of IL-17 and IFN-γ-producing T cells demonstrated byintracellular staining.

FIG. 13 shows KD025 activates STAT5 phosphorylation. Freshly purifiedCD4⁺ T cells were cultured for 2 days with stimulatory antibodiesagainst CD3/CD28 (5 μg/ml), TGF-β (5 ng/ml), IL-1β (50 ng/ml), and theindicated doses of the selective ROCK2 inhibitor KD025.

FIG. 14 shows Foxp3 expression in human CD4⁺ T cells treated with theindicated doses of the selective ROCK2 inhibitor KD025.

FIG. 15 shows the effect of KD025-mediated ROCK2 inhibition in Tregs onIL-17 secretion by CD4⁺CD25⁻ T cells.

FIG. 16 shows the effect of KD025-mediated ROCK2 inhibition onTGF-3-induced phosphorylation of STAT3, MLC, and SMAD2/3 in Tregs.

FIG. 17A-C shows the effect of KD025-mediated ROCK2 inhibition onstimulation of (A) IL-17, (B) IL-21, and (C) IFN-γ production inisolated PBMCs. Six patients were treated with 120 mg/day KD025 on days1 and 8-14, and two patients with placebo. PBMCs were isolated at days 1and 15 and examined for IL-17 and IL-21 production in response tostimulation by anti-CD3/CD28 antibodies. Patients 2 and 7 receivedplacebo. *=placebo administered to patients 2 and 7. Panels D-F show theeffect of increasing doses of KD025 on stimulation of (D) IL-17, (E)IL-21, and (F) IFN-γ production in isolated PBMCs.

FIG. 18 shows the effect of KD025-mediated ROCK2 inhibition on lungpathology in a mouse model of chronic GVHD. Panel A shows the results ofpulmonary function tests of volume, resistance, elastance, andcompliance in mice transplanted with bone marrow (BM) cells with orwithout allogeneic splenocytes (“vehicle”), and treated or untreatedwith KD025. Panels B and C show that body weight and survival ratesbetween groups of test animals were comparable, thus indicating thatKD025 has no effect on body weight and survival rates.

FIG. 19, Panel A, shows KD025 dose dependence of reversal ofcGVHD-associated lung pathology. Mice transplanted with bone marrow (BM)cells and allogeneic splenocytes (“vehicle”) were treated with 30 mg/kg,100 mg/kg, or 150 mg/kg of KD025 and evaluated for pulmonary resistance,elastance, and compliance. Panel B shows that body weights betweengroups of test animals were comparable, thus indicating that KD025 hasno effect on body weight.

FIG. 20 shows collagen deposition and antibody deposition in miceresulting from transplantation with donor bone marrow (BM) cells,induction with allogeneic splenocytes (“vehicle”), and KD025-mediatedROCK2 inhibition.

FIG. 21 shows germinal center (GC) formation and T helper cell (Tfh)activity in mice induced to chronic GVHD and reduction of GC activity byKD025-mediated ROCK2 inhibition.

FIG. 22 shows that treatment with KD025 from days 28-56 down-regulatedSTAT3 phosphorylation and up-regulated STAT5 phosphorylation. Thiseffect was followed by dose-dependent reduction in protein levels ofRORγt, IRF4 and Bcl6 transcription factors in spleens.

FIG. 23 shows that body weights between groups of test animals werecomparable, thus indicating that KD025 has no effect on body weight.

FIG. 24 shows a pharmacokinetic analysis of KD025 in murine chronicGVHD, demonstrating that KD025 is detectable in the blood of treatedmice and in a dose depended fashion.

FIG. 25 shows the effect of KD025-mediated ROCK2 inhibition on lungpathology in a mouse model of chronic GVHD. Partial improvement in therespiratory function was detectable 42 days after the lastadministration of KD025 in mice with cGVHD.

FIG. 26 shows that KD025 significantly decreases the GVHD score in amouse model for sclerodermatous cGVHD.

FIG. 27 shows an illustration of a sclerodermatous cGVHD mouse model.

FIG. 28 shows data indicating role of CD4+ T cells expressing STAT3 inmanifestation of scleroderma.

FIG. 29 shows reduced Th1 and Th17 cell numbers in a mouse model ofsclerodermatous chronic GVHD receiving CD4+ STAT3 KO T cells.

FIG. 30 shows increased amount of Tregs in a mouse model ofsclerodermatous chronic GVHD receiving CD4+ STAT3 KO T cells.

FIG. 31 shows that KD025 treatment blocks progression of sclerodermatouschronic GVHD in mice.

FIG. 32 shows decreased cervical LN T Cell IFNg in KD025 treatedscleroderma mice.

FIG. 33 shows IFNg in donor T Cells within the cervical lymph node (CLN)in KD025 treated scleroderma mice with chronic GVHD.

FIG. 34 shows the analysis of inflammation in KD025 treated sclerodermamice.

FIG. 35 shows macrophage infiltration and scleroderma in lethallyirradiated recipients of G-CSF mobilized fully allogeneic splenocytes ornon-mobilized semi-allogeneic splenocytes.

DETAILED DESCRIPTION

The present invention relates to compounds having the formula I:

wherein:

-   -   X is selected from N or C—R₁;    -   Y is selected from N or C—R₅;    -   Z is selected from N or C—R₃;    -   R₁ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₂ is a group having the formula -A-R₁₀;    -   A is selected from the group consisting of a covalent bond,        aryl, heteroaryl, cycloalkyl, and heterocyclyl;    -   R₁₀ is selected from the group consisting of H, CN, halo,        hydroxy, lower alkoxy, amino, perfluoro lower alkyl, C₁-C₁₀        alkyl, C₂-C₁₀ alkenyl, and -(M)_(x)-(CH₂)_(y)—R₁₁    -   M is selected from the group consisting of N—R₂₀, CR₂₁R₂₂, and        C═O;    -   x is 0 or 1;    -   R₂₀ is selected from H and C₁₋₅ alkyl;    -   R₂₁ and R₂₂ are independently selected from the group consisting        of H, halogen, and lower alkyl, or alternatively R₂₁ and R₂₂ may        be taken together with the atom to which they are attached to        form a C₃₋₆ cycloalkyl;    -   y is 0, 1, 2, 3, 4, 5, or 6;    -   R₁₁ is selected from the group consisting of H, C₁₋₆ alkyl,        optionally substituted C₃₋₆ cycloalkyl, optionally substituted        aryl, optionally substituted heteroaryl, optionally substituted        heterocyclyl, wherein the optional substituents are selected        from the group consisting of lower alkyl, C₁₋₆ cycloalkyl, oxo,        CN, halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl,        and (lower alkyl)-O-(lower alkyl);    -   alternatively R₁₁ is selected from the group consisting of        —NR₁₃R₁₄, —C(═O)NR₁₃R₁₄, and —C(═O)R₁₂, and —CO₂R₁₂;    -   R¹² is selected from the group consisting of C₁-C₁₀ alkyl, aryl,        heteroaryl, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), aralkyl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, oxo, C₁-C₆ alkyl, C₃-C₆ cycloalkyl, C₁-C₆ alkoxy,        (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), hydroxy, cyano and C₁-C₃        perfluoro alkyl;    -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₆        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, oxo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl,        C₁-C₆ alkoxy, (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), hydroxy, amino,        cyano and C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, (C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), oxo, hydroxy,        cyano and C₁-C₃ perfluoroalkyl;    -   R₃ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₅ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₆ is selected from the group consisting of H, lower alkyl, CN,        halo, hydroxy, lower alkoxy, amino, perfluoro lower alkyl, and        (lower alkyl)-O-(lower alkyl);    -   R₇ is selected from the group consisting of H and lower alkyl;        and    -   R₈ is a nitrogen-containing heterocyclic ring system ring which        may comprise 0-2 additional ring heteroatoms selected from N, O        and S, and may be unsubstituted or may be substituted with 1 to        3 substituents selected from halo, CN, oxo, hydroxy, amino,        lower alkyl, perfluoro lower alkyl, and lower alkoxy.

In certain embodiments of the invention, the ring system of R₈ issaturated, contains one or more double bonds, or is aromatic. The ringsystem than comprises R₈ is preferably a monocyclic or a bicyclic ringsystem having 4 to 10 ring atoms. In certain aspects of the invention,R₈ is selected from:

wherein R₉ is selected from H, halogen and lower alkyl.

In certain embodiments, R₂ is a substituted aryl group and is preferablya substituted phenyl group.

In certain aspects of the invention, the compounds useful according tothe present invention include those having the formula II, III or IV:

wherein R₂, R₆, R₇, X and Z are as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula V or VI:

wherein R₆, R₇, X, Z and R₁₀ are as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula VII:

wherein R₆, R₇, X, Z, and R₁₀ are as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula IX:

wherein R₆, R₇, X and Z are as defined above for formula I, and T is—(CH₂)_(y)—R₁₁ wherein y and R₁₁ are as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula X:

wherein R₆, R₇, X and Z are as defined above for formula I, and R′ isR₁₃ as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XI:

wherein R₆, R₇, X and Z are as defined above for formula I, and T₁ isR₁₂ as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XII:

wherein R₆, R₇, X and Z are as defined above for formula I, and T₁ isR₁₂ as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XIII:

wherein R₆, R₇, X and Z are as defined above for formula I, A is M asdefined above for formula 1 and W is R₁₂ as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XIV:

wherein X, Z and R¹³ are as defined above for formula I.

In certain aspects of the invention, for each of the compounds depictedabove, the moiety

may be selected from a heteroaromatic group such that Y is N. In otheraspects of the invention, both Y and X are N, and in still other aspectsX, Y and Z are each N. In preferred aspects of the invention, thisheteroaromatic group is selected from any one of the following groups:

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XV:

wherein R₆, R₇, and R₁₀ are as defined above for formula I.

In other aspects of the invention, the compounds useful according to thepresent invention include those having the formula XVI:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        R² is selected from H and halo;        each R³ and R⁴ is independently selected from the group        consisting of H, C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl),        —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², C₁-C₃ perfluoro        alkyl, —O—(CH₂)_(a)NR³¹R³², —NR³¹—(CH₂)_(a)NR³³R³⁴,        —NR³¹—(CH₂)_(a)OR³³, aryl, C₃-C₇ cycloalkyl, a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, and —(C₁-C₆ alkyl)-O—(C₁-C₆        alkyl);    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   R³³ and R³⁴ are independently selected from the group consisting        of H and C₁-C₈ alkyl;    -   a is selected from 0 to 6;        R⁵ is selected from H and C₁-C₆ alkyl;        R⁶ is selected from the group consisting of H, halo, and C₁-C₆        alkyl.

In an embodiment of the invention R¹³ is selected from the groupconsisting of C₁-C₈ alkyl, C₃-C₇ cycloalkyl and a three totwelve-membered heterocyclic ring. In an another embodiment of theinvention R¹³ is selected from the group consisting of isopropyl,cycloalkyl, N-morpholino and 3-pyridine. In an embodiment of theinvention R¹⁴ is H. In an embodiment of the invention R² is H. Inanother embodiment of the invention R² is F. In an embodiment of theinvention R³ is selected from the group consisting of H, C₁-C₈ alkyl andC₁-C₃ perfluoro alkyl. In an another embodiment of the invention R³ isselected from the group consisting of H, CH₃ and CF₃. In an embodimentof the invention R⁴ is selected from the group consisting of H, C₁-C₈alkyl, C₁-C₃ perfluoro alkyl and a three to twelve-membered heterocyclicring. In an another embodiment of the invention R⁴ is selected from thegroup consisting of H, CH₃, CF₃, piperazinyl and N-morpholino.

In aspects of the invention, the compounds useful according to thepresent invention include those having the formula XVII:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:X is selected from the group consisting of —NH—C(═O)—CHR¹³R¹⁴;—NH—C(═O)—(CH₂)_(b)—NR¹³R¹⁴; —C(═O)—NR¹³R¹⁴;

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), aryl, heteroaryl, C₃-C₇ cycloalkyl, a        three to twelve membered heterocyclic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;        each R³ and R⁴ is independently selected from the group        consisting of H, C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl),        —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², C₁-C₃ perfluoro        alkyl, —O—(CH₂)_(a)NR³¹R³², aryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic ring having up to 3 heteroatoms        which is optionally substituted from 1 to 3 substituents        independently selected from halo and C₁-C₆ alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   a is selected from 0 to 6;    -   b is selected from 0 to 1.

In an embodiment of the invention R¹³ is a three to twelve-memberedheterocyclic ring. In an another embodiment of the invention R¹³ isselected from the group consisting of isopropyl, cycloalkyl,N-morpholino, 3-pyridinyl, tetrahydropyranyl, piperdinyl, andtetrahydrothiopyranyl dioxide. In an another embodiment of the inventionR¹³ is selected from the group consisting of:

In an embodiment of the invention R¹⁴ is H. In an embodiment of theinvention R³ and R⁴ are each H.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XVIII:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:each R³ and R⁴ is independently selected from the group consisting of H,C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), —NR³¹R³², CF₃, —O—(CH₂)_(a)NR³¹R³², aryl, C₃-C₇ cycloalkyl, athree to twelve membered heterocyclic ring having up to 3 heteroatomswhich is optionally substituted from 1 to 3 substituents independentlyselected from halo and C₁-C₆ alkyl;

-   -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   a is selected from 0 to 6;        R¹⁵ is selected from the group consisting of H, C₁-C₈ alkyl,        —CN, halo, —OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆        alkyl), —C(═O)—O—C(R)₃ ³¹, CF₃, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo and C₁-C₆ alkyl;    -   x is selected from 1 to 3;    -   y is selected from 0 to 3;    -   z is selected from 0 to 3;    -   wherein y or z are independently selected and one of which is at        least 1.

In aspects of the invention, the compounds useful according to thepresent invention include those having the formula XIX:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        Y is selected from the group consisting of S, CH₂, and —CR³¹R³²—        R² is selected from H and halo;        each R³ and R⁴ is independently selected from the group        consisting of H, C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl),        —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², C₁-C₃ perfluoro        alkyl, —O—(CH₂)_(a)NR³¹R³², —NR³¹—(CH₂)_(a)NR³³R³⁴,        —NR³¹—(CH₂)_(a)OR³³, aryl, C₃-C₇ cycloalkyl, a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, and —(C₁-C₆ alkyl)-O—(C₁-C₆        alkyl);    -   R³¹ and R³² are independently selected from the group consisting        of H, halo, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and        —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered cycloalkyl or heterocyclic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo and C₁-C₆ alkyl;    -   R³³ and R³⁴ are independently selected from the group consisting        of H and C₁-C₈ alkyl;    -   a is selected from 0 to 6.

In an embodiment of the invention Y forms a three-membered cycloalkane.In an another embodiment of the invention Y is fluoro;

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XX:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        R⁴ is selected from the group consisting of H, C₁-C₈ alkyl, —CN,        halo, —OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),        —NR³¹R³², CF₃, —O—(CH₂)_(a)NR³¹R³², aryl, C₃-C₇ cycloalkyl, a        three to twelve membered heterocyclic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo and C₁-C₆ alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;        R⁵ is selected from H and C₁-C₆ alkyl.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XXI:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        R⁴ is selected from the group consisting of H, C₁-C₈ alkyl, —CN,        halo, —OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),        —NR³¹R³², C₁-C₃ perfluoro alkyl, —O—(CH₂)_(a)NR³¹R³², aryl,        C₃-C₇ cycloalkyl, a three to twelve membered heterocyclic ring        having up to 3 heteroatoms which is optionally substituted from        1 to 3 substituents independently selected from halo and C₁-C₆        alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   a is selected from 0 to 6.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XXII:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        R³ is H;        R⁴ is selected from the group consisting of H, C₁-C₈ alkyl, —CN,        halo, —OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),        —NR³¹R³², C₁-C₃ perfluoro alkyl, —O—(CH₂)_(a)NR³¹R³², aryl,        C₃-C₇ cycloalkyl, a three to twelve membered heterocyclic ring        having up to 3 heteroatoms which is optionally substituted from        1 to 3 substituents independently selected from halo and C₁-C₆        alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   a is selected from 0 to 6.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XXIII:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;        x is selected from 0 to 1;        R² is selected from the group consisting of cyclohexylpyridine,        1H-pyrazole, and pyridine;

X is selected from N or CR³;

Y is selected from N or CR³;

Z is selected from N or CR⁴;

wherein at least one of X, Y, and Z is N;

R⁴ is selected from the group consisting of H, C₁-C₈ alkyl, —CN, halo,—OH, —O—(C₁-C₆ alkyl), —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², CF₃,—O—(CH₂)_(a)NR³¹R³², —NR³¹—(CH₂)_(a)NR³³R³⁴, —NR³¹—(CH₂)_(a)OR³³, aryl,C₃-C₇ cycloalkyl, a three to twelve membered heterocyclic ring having upto 3 heteroatoms which is optionally substituted from 1 to 3substituents independently selected from halo, C₁-C₆ alkyl, and —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl);

-   -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   a is selected from 0 to 6;        Q is selected from the group NR⁵ and 0;        R⁵ is selected from H and C₁-C₆ alkyl.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XXIV:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹² is selected from the group consisting of H, C₁-C₈ alkyl,        C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),        amino, NR³¹R³², heteroaryl, C₃-C₇ cycloalkyl, a three to twelve        membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;        x is selected from 0 to 2;        each R³ and R⁴ is independently selected from the group        consisting of H, C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl),        —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², CF₃,        —O—(CH₂)_(a)NR³¹R³², aryl, C₃-C₇ cycloalkyl, a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₃-C₇        cycloalkyl and —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   a is selected from 1 to 6.

In another aspect of the invention, the compounds useful according tothe present invention include those having the formula XXV:

or pharmaceutically acceptable salt or stereoisomer thereof, wherein:

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), heteroaryl, C₃-C₇ cycloalkyl, a three to        twelve membered heterocyclic or aromatic ring containing up to 3        heteroatoms, each of which may be optionally substituted by from        1 to 3 substituents independently selected from halo, oxo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆ alkoxy, CN and        C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, —OH, —NH₂, CN and C₁-C₃ perfluoro alkyl;    -   x is selected from 0 to 3;        R¹⁵ is selected from the group consisting of H, C₁-C₈ alkyl,        C₂-C₈ alkenyl, C₂-C₈ alkynyl, aryl, heteroaryl, heterocyclic        ring, and C₃-C₇ cycloalkyl;        each R³ and R⁴ is independently selected from the group        consisting of H, C₁-C₈ alkyl, —CN, halo, —OH, —O—(C₁-C₆ alkyl),        —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², CF₃,        —O—(CH₂)_(a)NR³¹R³², aryl, C₃-C₇ cycloalkyl, a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        or R³¹ and R³² may be taken together to form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted from 1 to 3 substituents independently        selected from halo and C₁-C₆ alkyl;

a is selected from 1 to 6.

The present invention relates to compounds having the formula XXVI:

or pharmaceutically acceptable salt thereof, wherein:R¹ is selected from the group consisting of H or C₁-C₆ alkyl;R² is selected from the group consisting of aryl, heteroaryl, C₃-C₇cycloalkyl, a three to twelve membered heterocyclic ring containing upto 3 heteroatoms, each of which may be optionally substituted from 1 to3 substituents independently selected from halo and C₁-C₆ alkyl;

-   -   X is selected from N or CR³;    -   Y is selected from N or CR³;    -   Z is selected from N or CR⁴;    -   W is selected from CR⁵;    -   wherein X, Y, and Z are independently selected and at least one        of which is N;        wherein each R³ is independently selected from the group        consisting of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl,        —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR³¹R³², and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   R³¹ and R³² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);        R⁴ and R⁵ are independently selected from the group consisting        of H, halo, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, C₁-C₆        alkoxy, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —NR⁴¹R⁴²,        —(CH₂)_(x)NR⁴¹R⁴² and —O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl;    -   R⁴¹ and R⁴² are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, and —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl);    -   or R⁴¹ and R⁴² may be taken together to form a three to twelve        membered cycloalkyl or heterocyclic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo, C₁-C₆ alkyl, and        C₁-C₆ alkoxy;    -   x is selected from 0 to 6;        or R⁴ and R⁵ may be taken together to form a three to twelve        membered heterocyclic or aromatic ring having up to 3        heteroatoms which is optionally substituted from 1 to 3        substituents independently selected from halo and C₁-C₆ alkyl        and C₁-C₆ alkoxy;    -   a is selected from 0 to 2;    -   b is selected from 0 to 2;    -   wherein a or b are independently selected and one of which is at        least 1        A is a three to twelve membered heterocyclic or aromatic ring        having up to 3 heteroatoms which is optionally substituted from        1 to 3 substituents independently selected from halo, C₁-C₆        alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, C₃-C₇ cycloalkyl, oxo, acyl,        —S—(C₁-C₆ alkyl), OH, NH₂, CN and C₁-C₃ perfluroalkyl.

In an embodiment of the invention, R¹ is H;

In an embodiment of the invention, R² is selected from indazole,cyclohexylpyridine, phenylpyridine, phenyl-1H-pyrazole,1H-pyrrolo[2,3-b]pyridine, 1H-pyrazole, pyridine, isoquinoline,quinoline and 1,3-thiazolyl pyridine;

In an embodiment of the invention, R² is selected from:

In an embodiment of the invention R² is selected from:

-   -   X, Z═N; W═CR³    -   X, Y═N; Z═CR⁴

In an embodiment of the invention R⁴ and R⁵ are each H;

In another embodiment R⁴ and R⁵ are each —CH₃;

In still another embodiment R⁴ and R⁵ are taken together to form afive-membered ring including, without limitation, dihydrofuran;

In other aspects of the invention, the ROCK2 inhibiting compound may beselected from the ROCK2 compounds disclosed in PCT/US2006/011271, filedMar. 27, 2006, which is incorporated herein in its entirety. Thus, theROCK2 inhibiting compound may have the formula XXXI:

or pharmaceutically acceptable salt, wherein:R₁ is selected from the group consisting of —O—(CH₂)_(y)—CO₂R¹²,—O—(CH₂)_(y)—C(═O)NR¹³R¹⁴, —O—(CH₂)_(y)-heteroaryl,—O—(CH₂)_(y)-cycloalkyl, —O—C(═O)—(CH₂)_(y)—NR¹³R¹⁴,—O—(CH₂)_(z)—NR¹³R¹⁴, —NH—C(═O)—(CH₂)—NR¹³R¹⁴, —NH—C(═O)—X—R¹⁵,—NH—(CH₂)_(y)—NR¹³R¹⁴;R¹² is selected from the group consisting of C₁-C₆ alkyl, —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆alkyl)-C(═O)NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted at one or more carbon atoms by from 1 to 3substituents independently selected from halo, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;

-   -   R¹³ and R¹⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆        alkyl)-C(═O)NR¹⁶R¹⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R¹³ and R¹⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, OH, NH₂, CN and C₁-C₃ perfluoro alkyl;    -   X is selected from a covalent bond, O, NH, and C₁-C₆ alkyl;    -   R¹⁵ is selected from the group consisting of heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, OH, NH₂, CN        and C₁-C₃ perfluoro alkyl;    -   or R¹⁵ is selected from —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆        alkyl)-NR¹⁶R¹⁷, —CO₂R₈, —O—(CH₂)_(x)—CO₂R¹⁸, and —C(═O)NR¹⁶R¹⁷;        -   R¹⁶ and R¹⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, OH, NH₂, CN and C₁-C₃ perfluoro            alkyl;        -   or R¹⁶ and R¹⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R¹⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;    -   x is selected from 0 to 6;    -   y is selected from 0 to 6;    -   z is selected from 2 to 6;        each R² is independently selected from the group consisting of        lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and        perfluoro lower alkyl;        each R³ is independently selected from the group consisting of        lower alkyl, CN, halo, hydroxy, lower alkoxy, amino, and        perfluoro lower alkyl;        R⁴ is selected from —(CH₂)_(a)—NR⁴³R⁴⁴, —Y—R⁴²,        —O—(CH₂)_(a)—CO₂R⁴², —O—(CH₂)_(a)—C(═O)NR⁴³R⁴⁴,        —O—(CH₂)_(a)-heteroaryl, —O—(CH₂)_(a)-cycloalkyl,        —O—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴, —O—(CH₂)_(c)—NR⁴³R⁴⁴,        —NH—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴, —NH—C(═O)—Y—R⁴⁵,        —NH—C(═O)—(CH₂)_(a)—NR⁴³R⁴⁴;    -   R⁴² is selected from the group consisting of C₁-C₆ alkyl,        —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆        alkyl)-C(═O)NR⁴⁶R⁴⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆        alkyl), each of which may be optionally substituted at one or        more carbon atoms by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and        C₁-C₃ perfluoro alkyl;    -   R⁴³ and R⁴⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆        alkyl)-C(═O)NR⁴⁶R⁴⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁴³ and R⁴⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   Y is selected from a covalent bond, O, NH, and C₁-C₆ alkyl;    -   R⁴⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —CO₂R⁴⁸,        —O—(CH₂)_(b)—CO₂R⁴⁸, and —C(═O)NR⁴⁶R⁴⁷,        -   R⁴⁶ and R⁴⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁴⁶ and R⁴⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁴⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁴⁶R⁴⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;    -   a is selected from 0 to 6;    -   b is selected from 0 to 6;    -   c is selected from 2 to 6;        R⁵ is selected from the group consisting of H, C₁-C₆ alkyl,        —(CH₂)_(d)—C(═O)—NR⁵³R⁵⁴, —C(═O)—(CH₂)_(d)—NR⁵³R⁵⁴,        —C(═O)—X—R⁵⁵, and —C(═O)—(CH₂)_(d)—NR⁵³R⁵⁴;    -   R⁵³ and R⁵⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —(C₁-C₆        alkyl)-C(═O)NR⁵⁶R⁵⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁵³ and R⁵⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro        alkyl;    -   R⁵⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —CO₂R⁵⁸,        —O—(CH₂)_(e)—CO₂R⁵⁸, and —C(═O)NR⁵⁶R⁵⁷,        -   R⁵⁶ and R⁵⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁵⁶ and R⁵⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁵⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁵⁶R⁵⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;        -   d is selected from 0 to 6;        -   e is selected from 0 to 6;            R⁶ is selected from the group consisting of H, C₁-C₆ alkyl,            —(CH₂)_(r)—C(═O)—NR⁶³R⁶⁴, —C(═O)—(CH₂)_(r)—NR⁶³R⁶⁴,            —C(═O)—X—R⁶⁵, and —C(═O)—(CH₂)_(r)—NR⁶³R⁶⁴;    -   R⁶³ and R⁶⁴ are independently selected from the group consisting        of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —(C₁-C₆        alkyl)-C(═O)NR⁶⁶R⁶⁷, aryl, aralkyl, heteroaryl, C₃-C₇        cycloalkyl, a three to twelve membered heterocyclic ring        containing up to 3 heteroatoms, each of which may be optionally        substituted by from 1 to 3 substituents independently selected        from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆        alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;    -   or R⁶³ and R⁶⁴ may be taken together form a three to twelve        membered heterocyclic ring having up to 3 heteroatoms which is        optionally substituted by from 1 to 3 substituents independently        selected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy,        C₃-C₇ cycloalkyl, oxo, hydroxy, amino, cyano and C₁-C₃ perfluoro        alkyl;    -   R⁶⁵ is selected from the group consisting of H, aryl, —(C₁-C₆        alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —CO₂R⁶⁸,        —O—(CH₂)_(s)—CO₂R⁶⁸, and —C(═O)NR⁶⁶R⁶⁷,        -   R⁶⁶ and R⁶⁷ independently selected from the group consisting            of H, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₁-C₈ alkynyl, —(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), aryl, aralkyl, heteroaryl, C₃-C₇            cycloalkyl, a three to twelve membered heterocyclic ring            containing up to 3 heteroatoms, each of which may be            optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   or R⁶⁶ and R⁶⁷ may be taken together form a three to twelve            membered heterocyclic ring having up to 3 heteroatoms which            is optionally substituted by from 1 to 3 substituents            independently selected from halo, C₁-C₆ alkyl, C₂-C₆            alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino, cyano and C₁-C₃            perfluoro alkyl;        -   R⁶⁸ is selected from the group consisting of H, aryl,            aralkyl, heteroaryl, C₁-C₆ alkyl, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl), —(C₁-C₆ alkyl)-NR⁶⁶R⁶⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆            alkyl)-O—(C₁-C₆ alkyl), each of which may be optionally            substituted by from 1 to 3 substituents independently            selected from halo, C₁-C₆ alkoxy, hydroxy, amino, cyano and            C₁-C₃ perfluoroalkyl;        -   r is selected from 0 to 6;        -   s is selected from 0 to 6;            n is selected from 0 to 4;            m is selected from 0 to 3; and            p is selected from 0 and 1.

In one embodiment of Formula XXXI, R⁴ and R⁵ are independently selectedfrom H and C₁-C₆ alkyl. In another embodiment, R⁴ and R⁵ are H.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXII:

or a pharmaceutically acceptable salt thereof, wherein R¹, R², R³, n andm are as for the compound of the formula I.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXIII:

or a pharmaceutically acceptable salt thereof, wherein R¹, R², R³, n andm are as for the compound of the formula I.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXIV:

or a pharmaceutically acceptable salt thereof, wherein:R¹³ and R¹⁴ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-C(═O)NR¹⁶R¹⁷, aryl,aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;or R¹³ and R¹⁴ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, C₃-C₇ cycloalkyl, oxo,hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;X is selected from a covalent bond, O, NH, and C₁-C₆ alkyl;R¹⁶ and R¹⁷ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelvemembered heterocyclic ring containing up to 3 heteroatoms, each of whichmay be optionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;or R¹⁶ and R¹⁷ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino,cyano and C₁-C₃ perfluoro alkyl;each R² is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;each R³ is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;n is selected from 0 to 4; andm is selected from 0 to 3.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXIV_(a):

or a pharmaceutically acceptable salt thereof, wherein:R¹³ and R¹⁴ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-C(═O)NR¹⁶R¹⁷, aryl,aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;or R¹³ and R¹⁴ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino,cyano and C₁-C₃ perfluoro alkyl;R¹⁶ and R¹⁷ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelvemembered heterocyclic ring containing up to 3 heteroatoms, each of whichmay be optionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;or R¹⁶ and R¹⁷ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino,cyano and C₁-C₃ perfluoro alkyl.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXV:

or a pharmaceutically acceptable salt thereof, wherein:R¹² is selected from the group consisting of C₁-C₆ alkyl, —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆alkyl)-C(═O)NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted at one or more carbon atoms by from 1 to 3substituents independently selected from halo, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;each R² is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;each R³ is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;n is selected from 0 to 4; andm is selected from 0 to 3.

In an embodiment of the invention, the compound of formula XXXI has theformula XXXV_(a):

or a pharmaceutically acceptable salt thereof, wherein:R¹² is selected from the group consisting of C₁-C₆ alkyl, —(C₁-C₆alkyl)-O—(C₁-C₆ alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆alkyl)-C(═O)NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl)-O—(C₁-C₆ alkyl),aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted at one or more carbon atoms by from 1 to 3substituents independently selected from halo, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl.

In another embodiment of the invention, the rho kinase inhibitor has theXXXVI:

or a pharmaceutically acceptable salt thereof, wherein:R¹³ and R¹⁴ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-C(═O)NR¹⁶R¹⁷, aryl,aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;or R¹³ and R¹⁴ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, C₃-C₇ cycloalkyl, oxo,hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;R¹⁶ and R¹⁷ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelvemembered heterocyclic ring containing up to 3 heteroatoms, each of whichmay be optionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;or R¹⁶ and R¹⁷ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino,cyano and C₁-C₃ perfluoro alkyl;each R² is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;each R³ is independently selected from the group consisting of loweralkyl, CN, halo, hydroxy, lower alkoxy, amino, and perfluoro loweralkyl;n is selected from 0 to 4; andm is selected from 0 to 3.

In an embodiment of the invention, the compound of formula XXXVI has theformula XXXVI_(a):

or a pharmaceutically acceptable salt thereof, wherein:R¹³ and R¹⁴ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), —(C₁-C₆ alkyl)-NR¹⁶R¹⁷, —(C₁-C₆ alkyl)-C(═O)NR¹⁶R¹⁷, aryl,aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelve memberedheterocyclic ring containing up to 3 heteroatoms, each of which may beoptionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₃-C₇ cycloalkyl, C₁-C₆alkoxy, hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;or R¹³ and R¹⁴ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, C₃-C₇ cycloalkyl, oxo,hydroxy, amino, cyano and C₁-C₃ perfluoro alkyl;R¹⁶ and R¹⁷ are independently selected from the group consisting of H,C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, —(C₁-C₆ alkyl)-O—(C₁-C₆alkyl), aryl, aralkyl, heteroaryl, C₃-C₇ cycloalkyl, a three to twelvemembered heterocyclic ring containing up to 3 heteroatoms, each of whichmay be optionally substituted by from 1 to 3 substituents independentlyselected from halo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, hydroxy,amino, cyano and C₁-C₃ perfluoro alkyl;or R¹⁶ and R¹⁷ may be taken together to form a three to twelve memberedheterocyclic ring having up to 3 heteroatoms which is optionallysubstituted by from 1 to 3 substituents independently selected fromhalo, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₁-C₆ alkoxy, oxo, hydroxy, amino,cyano and C₁-C₃ perfluoro alkyl.

In further aspects of the invention, the compound of formula XXXI isSLx-2119:

In further aspects of the invention, the rho kinase inhibitor isselected from the group consisting of:

-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-isopropylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-(2-methoxyethyl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-(pyridin-3-yl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-1-(4-methylpiperazin-1-yl)ethanone,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-1-morpholinoethanone,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-methylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N—((R)-pyrrolidin-3-yl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N—((S)-pyrrolidin-3-yl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N—((R)-tetrahydrofuran-3-yl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-1-(piperidin-1-yl)ethanone,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-tert-butylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-ethylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-(cyanomethyl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-cyclobutylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-isobutylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-(2,2,2-trifluoroethyl)acetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-cyclohexylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-neopentylacetamide,-   2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenoxy)-N-(prop-2-ynyl)acetamide,-   N-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-4-methylpiperazine-1-carboxamide,-   3-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-1,1-dimethylurea,-   N-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-2-methoxyacetamide,-   methyl    2-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenylamino)-2-oxoacetate,-   1-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-3-(2-(dimethylamino)ethyl)urea,-   N-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-2-morpholinoacetamide,-   N-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)-3-(4-isopropylpiperazin-1-yl)propanamide,-   N-(3-(4-(1H-indazol-5-ylamino)quinazolin-2-yl)phenyl)piperidine-4-carboxamide,    and    N-(3-(4-(1H-indazol-5-ylamino)-6-(2-methoxyethoxy)quinazolin-2-yl)phenyl)butyramide.

In further aspects of the invention, the rho kinase inhibitor is

The term “heteroatom” as used herein means an atom of any element otherthan carbon or hydrogen. Preferred heteroatoms are nitrogen, oxygen, andsulfur.

The term “alkyl” refers to the radical of saturated aliphatic groups,including straight-chain alkyl groups and branched-chain alkyl groups.In preferred embodiments, a straight chain or branched chain alkyl has10 or fewer carbon atoms in its backbone (e.g., C₁-C₁₀ for straightchain, C₃-C₁₀ for branched chain). Likewise, preferred cycloalkyls havefrom 3-10 carbon atoms in their ring structure, and more preferably have3 to 6 carbons in the ring structure.

Unless the number of carbons is otherwise specified, “lower alkyl” asused herein means an alkyl group, as defined above, but having from oneto six carbons, and more preferably from one to four carbon atoms.Likewise, “lower alkenyl” and “lower alkynyl” have similar chain lengths(C₂-C₆). Preferred alkyl groups are lower alkyls. In preferredembodiments, a substituent designated herein as alkyl is a lower alkyl.

The term “cycloalkyl” refers to saturated, carbocyclic groups havingfrom 3 to 7 carbons in the ring. Preferred cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term “aralkyl”, as used herein, refers to an alkyl group substitutedwith an aryl group (e.g., an aromatic or heteroaromatic group).

The terms “alkenyl” and “alkynyl” refer to unsaturated aliphatic groupsanalogous in length and possible substitution to the alkyls describedabove, but that contain at least one double or triple bond respectively.

The term “aryl” as used herein includes 5- and 6-membered single-ringaromatic groups that may include from zero to four heteroatoms, forexample, benzene, pyrene, pyrrole, furan, thiophene, imidazole, oxazole,thiazole, triazole, pyrazole, pyridine, pyrazine, pyridazine andpyrimidine, and the like. Those aryl groups having heteroatoms in thering structure may also be referred to as “aryl heterocycles”,“heteroaromatics” or “heteroaryl”. The aromatic ring can be substitutedat one or more ring positions with such substituents as described above,for example, halogen, azide, alkyl, aralkyl, alkenyl, alkynyl,cycloalkyl, hydroxyl, alkoxyl, amino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, sulfonamido, ketone, aldehyde, ester, heterocyclyl, aromaticor heteroaromatic moieties, —CF₃, —CN, or the like. The term “aryl” alsoincludes polycyclic ring systems having two or more cyclic rings inwhich two or more carbons are common to two adjoining rings (the ringsare “fused rings”) wherein at least one of the rings is aromatic, e.g.,the other cyclic rings can be cycloalkyls, cycloalkenyls, aryls and/orheterocyclic groups.

The terms “heterocyclyl” or “heterocyclic group” refer to 3- to10-membered ring structures, more preferably 5- or 6-membered rings,whose ring structures include one to four heteroatoms. Heterocycles canalso be polycycles. Heterocyclic groups include, for example, thiophene,thianthrene, furan, pyran, isobenzofuran, chromene, xanthene,phenoxathiin, pyrrole, imidazole, pyrazole, isothiazole, isoxazole,pyridine, pyrazine, pyrimidine, pyridazine, indolizine, isoindole,indole, indazole, purine, quinolizine, isoquinoline, quinoline,phthalazine, naphthyridine, quinoxaline, quinazoline, cinnoline,pteridine, carbazole, carboline, phenanthridine, acridine, pyrimidine,phenanthroline, phenazine, phenarsazine, phenothiazine, furazan,phenoxazine, pyrrolidine, oxolane, thiolane, oxazole, piperidine,piperazine, morpholine, lactones, lactams such as azetidinones andpyrrolidinones, sultams, sultones, and the like. The heterocyclic ringcan be substituted at one or more positions with such substituents asdescribed above, as for example, halogen, alkyl, aralkyl, alkenyl,alkynyl, cycloalkyl, hydroxyl, amino, nitro, sulfhydryl, imino, amido,phosphonate, phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio,sulfonyl, ketone, aldehyde, ester, a heterocyclyl, an aromatic orheteroaromatic moiety, —CF₃, —CN, or the like.

The terms “polycyclyl” or “polycyclic group” refer to two or more rings(e.g., cycloalkyls, cycloalkenyls, aryls and/or heterocyclyls) in whichtwo or more carbons are common to two adjoining rings, e.g., the ringsare “fused rings”. Rings that are joined through non-adjacent atoms aretermed “bridged” rings. Each of the rings of the polycyclic group can besubstituted with such substituents as described above, for example,halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, amino,nitro, sulfhydryl, imino, amido, phosphonate, phosphinate, carbonyl,carboxyl, silyl, ether, alkylthio, sulfonyl, ketone, aldehyde, ester, aheterocyclyl, an aromatic or heteroaromatic moiety, —CF₃, —CN, or thelike.

As used herein, the term “nitro” means —NO₂. The term “halogen” or“halo” designates —F, —Cl, —Br or —I. The term “hydroxyl” means —OH.

The terms “amine” and “amino” refer to both unsubstituted andsubstituted amines, e.g., a moiety that can be represented by thegeneral formula:

wherein R, R′ and R″ each independently represent H, alkyl, alkenyl,alkynyl, aralkyl, aryl, and heterocyclic groups, and most preferably Hor lower alkyl.

The terms “alkoxyl” or “alkoxy” as used herein refers to an alkyl group,as defined above, having an oxygen radical attached thereto.Representative alkoxyl groups include methoxy, ethoxy, propyloxy,tert-butoxy and the like. The term lower alkoxy refers to an alkoxygroup having from 1 to 6 carbon atoms.

The term “oxo” as used herein refers to an oxygen atom that has a doublebond to a another atom, particularly to carbon or sulfur.

As used herein, the definition of each expression, e.g. alkyl, m, n, R,etc., when it occurs more than once in any structure, is intended to beindependent of its definition elsewhere in the same structure.

It will be understood that “substituted”, “substitution” or “substitutedwith” includes the implicit proviso that such substitution is inaccordance with permitted valence of the substituted atom and thesubstituent, and that the substitution results in a stable compound,e.g., which does not spontaneously undergo transformation such as byrearrangement, cyclization, elimination, etc.

As used herein, the term “substituted” is contemplated to include allpermissible substituents of organic compounds. In a broad aspect, thepermissible substituents include acyclic and cyclic, branched andunbranched, carbocyclic and heterocyclic, aromatic and non-aromaticsubstituents of organic compounds. Illustrative substituents include,for example, those described herein above.

Certain compounds of the present invention may exist in particulargeometric or stereoisomeric forms. The present invention contemplatesall such compounds, including cis- and trans-isomers, R- andS-enantiomers, diastereomers, the racemic mixtures thereof, and othermixtures thereof, as falling within the scope of the invention.Additional asymmetric carbon atoms may be present in a substituent suchas an alkyl group. All such isomers, as well as mixtures thereof, areincluded in this invention.

Certain embodiments of the present compounds may contain a basicfunctional group, such as amino or alkylamino, and are, thus, capable offorming pharmaceutically-acceptable salts withpharmaceutically-acceptable acids. The term “pharmaceutically-acceptablesalts” in this context, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.Representative salts include the hydrochloride, hydrobromide, sulfate,bisulfate, phosphate, nitrate, acetate, stearate, laurate, benzoate,lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate,tartrate, napthylate, and mesylate salts and the like. (See, forexample, Berge et al. “Pharmaceutical Salts”, J. Pharm. Sci. (1977)66:1-19).

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically-acceptable salts with pharmaceutically-acceptablebases. Representative salts include alkali or alkaline earth salts suchas lithium, sodium, potassium, calcium, magnesium salts and the like.Representative organic amines useful for the formation of base additionsalts include ethylamine, diethylamine, ethylenediamine, ethanolamine,diethanolamine, piperazine and the like. (See, for example, Berge etal., supra).

In one aspect, the present invention provides compounds of Formula Ithat are inhibitors of Rho-kinase. Rho kinase (ROCK), a serine/threoninekinase, serves as a target protein for small GTP-binding protein Rho,and is an important mediator of numerous cellular functions, includingfocal adhesions, motility, smooth muscle contraction, and cytokinesis.In smooth muscle, ROCK plays an important role in Ca²⁺ sensitization andthe control of vascular tone. It modulates the level of phosphorylationof the myosin II light chain of myosin II, mainly through inhibition ofmyosin phosphatase, and contributes to agonist-induced Ca²⁺sensitization in smooth muscle contraction.

Rho kinase is found in two forms, ROCK 1 (ROCKβ; p160-ROCK) and ROCK 2(ROCKα). In some embodiments, the compound of Formula I is selectivelyinhibits ROCK1. In some embodiments, the compound of Formula Iselectively inhibits ROCK2. In some embodiments, the compound of FormulaI is non-selective with respect to inhibition of ROCK1 and ROCK2.

Methods of determining kinase inhibition are well known in the art. Forexample, kinase activity of an enzyme and the inhibitory capacity of atest compound can be determined by measuring enzyme specificphosphorylation of a substrate. Commercial assays and kits can beemployed. For example, kinase inhibition can be determined using anIMAP® assay (Molecular Devices). This assay method involves the use of afluorescently-tagged peptide substrate. Phosphorylation of the taggedpeptide by a kinase of interest promotes binding of the peptide to atrivalent metal-based nanoparticle via the specific, high affinityinteraction between the phospho-group and the trivalent metal. Proximityto the nanoparticle results in increased fluorescence polarization.Inhibition of the kinase by a kinase inhibitor prevents phosphorylationof the substrate and thereby limits binding of the fluorescently-taggedsubstrate to the nanoparticle. Such an assay can be compatible with amicrowell assay format, allowing simultaneous determination of IC₅₀ ofmultiple compounds.

In another aspect of the present invention there is provided a method oftreating a patient suffering from a disease comprising administering toa patient in need of such treatment a therapeutically effective amountof a compound of the present invention. The phrase“therapeutically-effective amount” as used herein means that amount of acompound, material, or composition comprising a compound of the presentinvention which is effective for producing some desired therapeuticeffect in at least a sub-population of cells in an animal at areasonable benefit/risk ratio applicable to any medical treatment, e.g.reasonable side effects applicable to any medical treatment.

Compounds of the invention that inhibit Rho-kinase and or Rho-kinasemediated phosphorylation are useful for treatment of patients sufferingfrom cardiovascular and non-cardiovascular diseases involving Rho-kinasefunction, such as hypertension, pulmonary hypertension, atherosclerosis,restenosis, coronary heart disease, cardiac hypertrophy, ocularhypertension, retinopathy, ischemic diseases, cerebral ischemia,cerebral vasospasm, penile erectile dysfunction, peripheral circulatorydisorder, peripheral artery occlusive disease, glaucoma, (e.g.,regulating intraoccular pressure), fibroid lung, fibroid liver, fibroidkidney, chronic obstructive pulmonary disease (COPD), adult respiratorydistress syndrome, central nervous system disorders such as neuronaldegeneration and spinal cord injury. Further, Rho-kinase inhibitors ofthe invention can be used to treat arterial thrombotic disorders such asplatelet aggregation and leukocyte aggregation, and bone resorption.

In certain embodiments, a Rho-kinase inhibitor of the invention is usedto treat inflammation, including, but not limited to asthma,cardiovascular inflammation, renal inflammation, and arteriosclerosis.

Rho-kinase inhibitors of the invention inhibit tumor cell growth andmetastasis, and angiogenesis, and are useful for treating neoplasticdiseases. Neoplastic diseases include any malignant growth or tumorcaused by abnormal or uncontrolled cell division, and may spread toother parts of the body through the lymphatic system or the bloodstream. Neoplastic disease includes, without limitation, lymphoma (aneoplasm of lymph tissue that is usually malignant), carcinoma (anymalignant tumor derived from epithelial tissue), leukemia (malignantneoplasm of blood-forming tissues; characterized by abnormalproliferation of leukocytes), sarcoma (a usually malignant tumor arisingfrom connective tissue (bone or muscle etc.), and blastoma (malignancyin precursor cells). Nonlimiting examples include squamous cell cancer,small-cell lung cancer, pituitary cancer, esophageal cancer,astrocytoma, soft tissue sarcoma, non-small cell lung cancer,adenocarcinoma of the lung, squamous carcinoma of the lung, cancer ofthe peritoneum, hepatocellular cancer, gastrointestinal cancer,pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, livercancer, bladder cancer, hepatoma, breast cancer, colon cancer,colorectal cancer, endometrial or uterine carcinoma, salivary glandcarcinoma, kidney cancer, liver cancer, prostate cancer, vulval cancer,thyroid cancer, hepatic carcinoma, brain cancer, endometrial cancer,testis cancer, cholangiocarcinoma, gallbladder carcinoma, gastriccancer, melanoma, and various types of head and neck cancer.

According to the invention, ROCK inhibitors are used to effect weightloss and/or limit weight gain. In a preferred embodiment, the ROCKinhibitor is ROCK2 selective. ROCK-2 inhibitors promote weight loss innormal subjects, and limit weight gain in subjects prone to obesity.

In an embodiment of the invention, a ROCK inhibitor is used to reduce orprevent insulin resistance or restore insulin sensitivity. Accordingly,in one embodiment, the compounds of the invention are used to promote orrestore insulin-dependent glucose uptake. In another embodiment of theinvention, a ROCK-inhibitors of the invention is used to promote orrestore glucose tolerance. In another embodiment of the invention, aROCK inhibitor of the invention is used to treat metabolic syndrome. Inanother embodiment, a ROCK-inhibitors of the invention is used to reduceor prevent hyperinsulinemia. In an embodiment of the invention, a ROCKinhibitor is used to treat diabetes (particularly type 2 diabetes). ROCKinhibitors of the invention may also be used to promote or restoreinsulin-mediated relaxation of vascular smooth muscle cells (VSMCs). Inpreferred embodiments, the ROCK inhibitor is ROCK2 selective.

In certain embodiments, compounds of the invention are used fortreatment of central nervous system disorders. Such disorders mayinvolve neuronal degeneration or physical injury to neural tissue,including without limitation, Huntington's disease, Parkinson's Disease,Alzheimer's, Amyotrophic lateral sclerosis (ALS), or multiple sclerosis.In certain embodiments, compounds of the invention have propertiesparticularly useful for treatment of such disorders, such as beneficialtissue distribution to tissues of the central nervous system, andability to cross the blood brain barrier.

The invention provides pan-ROCK inhibitors (i.e., compounds that inhibitROCK1 and ROCK1) as well as ROCK inhibitors that are isoform selective.As discussed above, in certain embodiments of the invention, aROCK2-selective inhibitor may be preferred. For example, one studyobserved that ROCK2 is frequently over expressed in hepatocellularcancer compared to non-timorous livers while ROCK1 expression isunaltered. Other cancers which may benefit from treatment with a ROCK2selective inhibitor include, but are not limited to, colon and bladdercancer. In contrast, ROCK1 expression levels have been observed to behigher in mammary tumors. Any cancer may be tested to determine whetherthere is overexpression of ROCK1 and/or ROCK2 and treated accordingly.In certain circumstances, ROCK 1 and ROCK2 isoforms show similarity inregulating certain downstream targets,

In another aspect, the present invention provides pharmaceuticallyacceptable compositions which comprise a therapeutically-effectiveamount of one or more of the compounds of Formula I, formulated togetherwith one or more pharmaceutically acceptable carriers (additives) and/ordiluents. As described in detail below, the pharmaceutical compositionsof the present invention may be specially formulated for administrationin solid or liquid form, including those adapted for the following: (1)oral administration, for example, drenches (aqueous or non-aqueoussolutions or suspensions), tablets, e.g., those targeted for buccal,sublingual, and systemic absorption, boluses, powders, granules, pastesfor application to the tongue; (2) parenteral administration, forexample, by subcutaneous, intramuscular, intravenous or epiduralinjection as, for example, a sterile solution or suspension, orsustained-release formulation; (3) topical application, for example, asa cream, ointment, or a controlled-release patch or spray applied to theskin; (4) intravaginally or intrarectally, for example, as a pessary,cream or foam; (5) sublingually; (6) ocularly; (7) transdermally; or (8)nasally.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals with toxicity, irritation,allergic response, or other problems or complications, commensurate witha reasonable benefit/risk ratio.

The phrase “pharmaceutically-acceptable carrier” as used herein means apharmaceutically-acceptable material, composition or vehicle, such as aliquid or solid filler, diluent, excipient, manufacturing aid (e.g.,lubricant, talc magnesium, calcium or zinc stearate, or steric acid), orsolvent encapsulating material, involved in carrying or transporting thesubject compound from one organ, or portion of the body, to anotherorgan, or portion of the body. Each carrier must be “acceptable” in thesense of being compatible with the other ingredients of the formulationand not injurious to the patient. Some examples of materials which canserve as pharmaceutically-acceptable carriers include: (1) sugars, suchas lactose, glucose and sucrose; (2) starches, such as corn starch andpotato starch; (3) cellulose, and its derivatives, such as sodiumcarboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4)powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients,such as cocoa butter and suppository waxes; (9) oils, such as peanutoil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; (10) glycols, such as propylene glycol; (11) polyols, suchas glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters,such as ethyl oleate and ethyl laurate; (13) agar; (14) bufferingagents, such as magnesium hydroxide and aluminum hydroxide; (15) alginicacid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer'ssolution; (19) ethyl alcohol; (20) pH buffered solutions; (21)polyesters, polycarbonates and/or polyanhydrides; and (22) othernon-toxic compatible substances employed in pharmaceutical formulations.

As set out above, certain embodiments of the present compounds maycontain a basic functional group, such as amino or alkylamino, and are,thus, capable of forming pharmaceutically-acceptable salts withpharmaceutically-acceptable acids. The term “pharmaceutically-acceptablesalts” in this respect, refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ in the administration vehicle or thedosage form manufacturing process, or by separately reacting a purifiedcompound of the invention in its free base form with a suitable organicor inorganic acid, and isolating the salt thus formed during subsequentpurification. Representative salts include the hydrobromide,hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate,valerate, oleate, palmitate, stearate, laurate, benzoate, lactate,phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate,napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonatesalts and the like. (See, for example, Berge et al. (1977)“Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19).

The pharmaceutically acceptable salts of the subject compounds includethe conventional nontoxic salts or quaternary ammonium salts of thecompounds, e.g., from non-toxic organic or inorganic acids. For example,such conventional nontoxic salts include those derived from inorganicacids such as hydrochloride, hydrobromic, sulfuric, sulfamic,phosphoric, nitric, and the like; and the salts prepared from organicacids such as acetic, propionic, succinic, glycolic, stearic, lactic,malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxymaleic,phenylacetic, glutamic, benzoic, salicyclic, sulfanilic,2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethanedisulfonic, oxalic, isothionic, and the like.

In other cases, the compounds of the present invention may contain oneor more acidic functional groups and, thus, are capable of formingpharmaceutically-acceptable salts with pharmaceutically-acceptablebases. The term “pharmaceutically-acceptable salts” in these instancesrefers to the relatively non-toxic, inorganic and organic base additionsalts of compounds of the present invention. These salts can likewise beprepared in situ in the administration vehicle or the dosage formmanufacturing process, or by separately reacting the purified compoundin its free acid form with a suitable base, such as the hydroxide,carbonate or bicarbonate of a pharmaceutically-acceptable metal cation,with ammonia, or with a pharmaceutically-acceptable organic primary,secondary or tertiary amine. Representative alkali or alkaline earthsalts include the lithium, sodium, potassium, calcium, magnesium, andaluminum salts and the like. Representative organic amines useful forthe formation of base addition salts include ethylamine, diethylamine,ethylenediamine, ethanolamine, diethanolamine, piperazine and the like.(See, for example, Berge et al., supra).

Wetting agents, emulsifiers and lubricants, such as sodium laurylsulfate and magnesium stearate, as well as coloring agents, releaseagents, coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the compositions.

Examples of pharmaceutically-acceptable antioxidants include: (1) watersoluble antioxidants, such as ascorbic acid, cysteine hydrochloride,sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2)oil-soluble antioxidants, such as ascorbyl palmitate, butylatedhydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propylgallate, alpha-tocopherol, and the like; and (3) metal chelating agents,such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol,tartaric acid, phosphoric acid, and the like.

Formulations of the present invention include those suitable for oral,nasal, topical (including buccal and sublingual), rectal, vaginal and/orparenteral administration. The formulations may conveniently bepresented in unit dosage form and may be prepared by any methods wellknown in the art of pharmacy. The amount of active ingredient which canbe combined with a carrier material to produce a single dosage form willvary depending upon the host being treated, the particular mode ofadministration. The amount of active ingredient which can be combinedwith a carrier material to produce a single dosage form will generallybe that amount of the compound which produces a therapeutic effect.Generally, out of one hundred percent, this amount will range from about0.1 percent to about ninety-nine percent of active ingredient,preferably from about 5 percent to about 70 percent, most preferablyfrom about 10 percent to about 30 percent.

In certain embodiments, a formulation of the present invention comprisesan excipient selected from the group consisting of cyclodextrins,celluloses, liposomes, micelle forming agents, e.g., bile acids, andpolymeric carriers, e.g., polyesters and polyanhydrides; and a compoundof the present invention. In certain embodiments, an aforementionedformulation renders orally bioavailable a compound of the presentinvention.

Methods of preparing these formulations or compositions include the stepof bringing into association a compound of the present invention withthe carrier and, optionally, one or more accessory ingredients. Ingeneral, the formulations are prepared by uniformly and intimatelybringing into association a compound of the present invention withliquid carriers, or finely divided solid carriers, or both, and then, ifnecessary, shaping the product.

Formulations of the invention suitable for oral administration may be inthe form of capsules, cachets, pills, tablets, lozenges (using aflavored basis, usually sucrose and acacia or tragacanth), powders,granules, or as a solution or a suspension in an aqueous or non-aqueousliquid, or as an oil-in-water or water-in-oil liquid emulsion, or as anelixir or syrup, or as pastilles (using an inert base, such as gelatinand glycerin, or sucrose and acacia) and/or as mouth washes and thelike, each containing a predetermined amount of a compound of thepresent invention as an active ingredient. A compound of the presentinvention may also be administered as a bolus, electuary or paste.

In solid dosage forms of the invention for oral administration(capsules, tablets, pills, dragees, powders, granules, trouches and thelike), the active ingredient is mixed with one or morepharmaceutically-acceptable carriers, such as sodium citrate ordicalcium phosphate, and/or any of the following: (1) fillers orextenders, such as starches, lactose, sucrose, glucose, mannitol, and/orsilicic acid; (2) binders, such as, for example, carboxymethylcellulose,alginates, gelatin, polyvinyl pyrrolidone, sucrose and/or acacia; (3)humectants, such as glycerol; (4) disintegrating agents, such asagar-agar, calcium carbonate, potato or tapioca starch, alginic acid,certain silicates, and sodium carbonate; (5) solution retarding agents,such as paraffin; (6) absorption accelerators, such as quaternaryammonium compounds and surfactants, such as poloxamer and sodium laurylsulfate; (7) wetting agents, such as, for example, cetyl alcohol,glycerol monostearate, and non-ionic surfactants; (8) absorbents, suchas kaolin and bentonite clay; (9) lubricants, such as talc, calciumstearate, magnesium stearate, solid polyethylene glycols, sodium laurylsulfate, zinc stearate, sodium stearate, stearic acid, and mixturesthereof; (10) coloring agents; and (11) controlled release agents suchas crospovidone or ethyl cellulose. In the case of capsules, tablets andpills, the pharmaceutical compositions may also comprise bufferingagents. Solid compositions of a similar type may also be employed asfillers in soft and hard-shelled gelatin capsules using such excipientsas lactose or milk sugars, as well as high molecular weight polyethyleneglycols and the like.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared usingbinder (for example, gelatin or hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (for example,sodium starch glycolate or cross-linked sodium carboxymethyl cellulose),surface-active or dispersing agent. Molded tablets may be made bymolding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

The tablets, and other solid dosage forms of the pharmaceuticalcompositions of the present invention, such as dragees, capsules, pillsand granules, may optionally be scored or prepared with coatings andshells, such as enteric coatings and other coatings well known in thepharmaceutical-formulating art. They may also be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropylmethyl cellulose in varying proportionsto provide the desired release profile, other polymer matrices,liposomes and/or microspheres. They may be formulated for rapid release,e.g., freeze-dried. They may be sterilized by, for example, filtrationthrough a bacteria-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedin sterile water, or some other sterile injectable medium immediatelybefore use. These compositions may also optionally contain opacifyingagents and may be of a composition that they release the activeingredient(s) only, or preferentially, in a certain portion of thegastrointestinal tract, optionally, in a delayed manner. Examples ofembedding compositions which can be used include polymeric substancesand waxes. The active ingredient can also be in micro-encapsulated form,if appropriate, with one or more of the above-described excipients.

Liquid dosage forms for oral administration of the compounds of theinvention include pharmaceutically acceptable emulsions, microemulsions,solutions, suspensions, syrups and elixirs. In addition to the activeingredient, the liquid dosage forms may contain inert diluents commonlyused in the art, such as, for example, water or other solvents,solubilizing agents and emulsifiers, such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor and sesame oils),glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acidesters of sorbitan, and mixtures thereof.

Besides inert diluents, the oral compositions can also include adjuvantssuch as wetting agents, emulsifying and suspending agents, sweetening,flavoring, coloring, perfuming and preservative agents.

Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar and tragacanth, and mixturesthereof.

Formulations of the pharmaceutical compositions of the invention forrectal or vaginal administration may be presented as a suppository,which may be prepared by mixing one or more compounds of the inventionwith one or more suitable nonirritating excipients or carrierscomprising, for example, cocoa butter, polyethylene glycol, asuppository wax or a salicylate, and which is solid at room temperature,but liquid at body temperature and, therefore, will melt in the rectumor vaginal cavity and release the active compound.

Formulations of the present invention which are suitable for vaginaladministration also include pessaries, tampons, creams, gels, pastes,foams or spray formulations containing such carriers as are known in theart to be appropriate.

Dosage forms for the topical or transdermal administration of a compoundof this invention include powders, sprays, ointments, pastes, creams,lotions, gels, solutions, patches and inhalants. The active compound maybe mixed under sterile conditions with a pharmaceutically-acceptablecarrier, and with any preservatives, buffers, or propellants which maybe required.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients, such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to a compound of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants, suchas chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons,such as butane and propane.

Transdermal patches have the added advantage of providing controlleddelivery of a compound of the present invention to the body. Such dosageforms can be made by dissolving or dispersing the compound in the propermedium. Absorption enhancers can also be used to increase the flux ofthe compound across the skin. The rate of such flux can be controlled byeither providing a rate controlling membrane or dispersing the compoundin a polymer matrix or gel.

Ophthalmic formulations, eye ointments, powders, solutions and the like,are also contemplated as being within the scope of this invention.

Pharmaceutical compositions of this invention suitable for parenteraladministration comprise one or more compounds of the invention incombination with one or more pharmaceutically-acceptable sterileisotonic aqueous or nonaqueous solutions, dispersions, suspensions oremulsions, or sterile powders which may be reconstituted into sterileinjectable solutions or dispersions just prior to use, which may containsugars, alcohols, antioxidants, buffers, bacteriostats, solutes whichrender the formulation isotonic with the blood of the intended recipientor suspending or thickening agents.

Examples of suitable aqueous and nonaqueous carriers which may beemployed in the pharmaceutical compositions of the invention includewater, ethanol, polyols (such as glycerol, propylene glycol,polyethylene glycol, and the like), and suitable mixtures thereof,vegetable oils, such as olive oil, and injectable organic esters, suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of coating materials, such as lecithin, by the maintenance of therequired particle size in the case of dispersions, and by the use ofsurfactants.

These compositions may also contain adjuvants such as preservatives,wetting agents, emulsifying agents and dispersing agents. Prevention ofthe action of microorganisms upon the subject compounds may be ensuredby the inclusion of various antibacterial and antifungal agents, forexample, paraben, chlorobutanol, phenol sorbic acid, and the like. Itmay also be desirable to include isotonic agents, such as sugars, sodiumchloride, and the like into the compositions. In addition, prolongedabsorption of the injectable pharmaceutical form may be brought about bythe inclusion of agents which delay absorption such as aluminummonostearate and gelatin.

In some cases, in order to prolong the effect of a drug, it is desirableto slow the absorption of the drug from subcutaneous or intramuscularinjection. This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material having poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally-administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle.

Injectable depot forms are made by forming microencapsule matrices ofthe subject compounds in biodegradable polymers such aspolylactide-polyglycolide. Depending on the ratio of drug to polymer,and the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions which are compatible with body tissue.

When the compounds of the present invention are administered aspharmaceuticals, to humans and animals, they can be given per se or as apharmaceutical composition containing, for example, 0.1 to 99% (morepreferably, 10 to 30%) of active ingredient in combination with apharmaceutically acceptable carrier.

The preparations of the present invention may be given orally,parenterally, topically, or rectally. They are of course given in formssuitable for each administration route. For example, they areadministered in tablets or capsule form, by injection, inhalation, eyelotion, ointment, suppository, etc. administration by injection,infusion or inhalation; topical by lotion or ointment; and rectal bysuppositories. Oral administrations are preferred.

The phrases “parenteral administration” and “administered parenterally”as used herein means modes of administration other than enteral andtopical administration, usually by injection, and includes, withoutlimitation, intravenous, intramuscular, intraarterial, intrathecal,intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal,transtracheal, subcutaneous, subcuticular, intraarticulare, subcapsular,subarachnoid, intraspinal and intrasternal injection and infusion.

The phrases “systemic administration,” “administered systemically,”“peripheral administration” and “administered peripherally” as usedherein mean the administration of a compound, drug or other materialother than directly into the central nervous system, such that it entersthe patient's system and, thus, is subject to metabolism and other likeprocesses, for example, subcutaneous administration.

These compounds may be administered to humans and other animals fortherapy by any suitable route of administration, including orally,nasally, as by, for example, a spray, rectally, intravaginally,parenterally, intracisternally and topically, as by powders, ointmentsor drops, including buccally and sublingually.

Regardless of the route of administration selected, the compounds of thepresent invention, which may be used in a suitable hydrated form, and/orthe pharmaceutical compositions of the present invention, are formulatedinto pharmaceutically-acceptable dosage forms by conventional methodsknown to those of skill in the art.

Actual dosage levels of the active ingredients in the pharmaceuticalcompositions of this invention may be varied so as to obtain an amountof the active ingredient which is effective to achieve the desiredtherapeutic response for a particular patient, composition, and mode ofadministration, without being toxic to the patient.

The selected dosage level will depend upon a variety of factorsincluding the activity of the particular compound of the presentinvention employed, or the ester, salt or amide thereof, the route ofadministration, the time of administration, the rate of excretion ormetabolism of the particular compound being employed, the rate andextent of absorption, the duration of the treatment, other drugs,compounds and/or materials used in combination with the particularcompound employed, the age, sex, weight, condition, general health andprior medical history of the patient being treated, and like factorswell known in the medical arts.

A physician or veterinarian having ordinary skill in the art can readilydetermine and prescribe the effective amount of the pharmaceuticalcomposition required. For example, the physician or veterinarian couldstart doses of the compounds of the invention employed in thepharmaceutical composition at levels lower than that required in orderto achieve the desired therapeutic effect and gradually increase thedosage until the desired effect is achieved.

In general, a suitable daily dose of a compound of the invention will bethat amount of the compound which is the lowest dose effective toproduce a therapeutic effect. Such an effective dose will generallydepend upon the factors described above. Generally, oral, intravenous,intracerebroventricular and subcutaneous doses of the compounds of thisinvention for a patient, when used for the indicated analgesic effects,will range from about 0.0001 to about 100 mg per kilogram of body weightper day.

In certain embodiments, a dose of a compound or a composition isadministered to a subject every day, every other day, every couple ofdays, every third day, once a week, twice a week, three times a week, oronce every two weeks. If desired, the effective daily dose of the activecompound may be administered as two, three, four, five, six or moresub-doses administered separately at appropriate intervals throughoutthe day, optionally, in unit dosage forms. In some embodiments, adose(s) of a compound or a composition is administered for 2 days, 3days, 5 days, 7 days, 14 days, or 21 days. In certain embodiments, adose of a compound or a composition is administered for 1 month, 1.5months, 2 months, 2.5 months, 3 months, 4 months, 5 months, 6 months ormore.

The above-described administration schedules are provided forillustrative purposes only and should not be considered limiting. Aperson of ordinary skill in the art will readily understand that alldoses are within the scope of the invention.

While it is possible for a compound of the present invention to beadministered alone, it is preferable to administer the compound as apharmaceutical formulation (composition).

The patient receiving this treatment is any animal in need, includingprimates, in particular humans, and other mammals such as equines,cattle, swine and sheep; and poultry and pets in general.

The compound of the invention can be administered as such or inadmixtures with pharmaceutically acceptable carriers and can also beadministered in conjunction with antimicrobial agents such aspenicillins, cephalosporins, aminoglycosides and glycopeptides.Conjunctive therapy, thus includes sequential, simultaneous and separateadministration of the active compound in a way that the therapeuticaleffects of the first administered one is not entirely disappeared whenthe subsequent is administered.

The addition of the active compound of the invention to animal feed ispreferably accomplished by preparing an appropriate feed premixcontaining the active compound in an effective amount and incorporatingthe premix into the complete ration.

Alternatively, an intermediate concentrate or feed supplement containingthe active ingredient can be blended into the feed. The way in whichsuch feed premixes and complete rations can be prepared and administeredare described in reference books (such as “Applied Animal Nutrition”,W.H. Freedman and CO., San Francisco, U.S.A., 1969 or “Livestock Feedsand Feeding” O and B books, Corvallis, Oreg., U.S.A., 1977).

Microemulsification technology may be employed to improvebioavailability of lipophilic (water insoluble) pharmaceutical agents.Examples include Trimetrine (Dordunoo, S. K., et al., Drug Developmentand Industrial Pharmacy, 17(12), 1685-1713, 1991) and REV 5901 (Sheen,P. C., et al., J Pharm Sci 80(7), 712-714, 1991). Among other things,microemulsification provides enhanced bioavailability by preferentiallydirecting absorption to the lymphatic system instead of the circulatorysystem, which thereby bypasses the liver, and prevents destruction ofthe compounds in the hepatobiliary circulation.

In one aspect of invention, the formulations contain micelles formedfrom a compound of the present invention and at least one amphiphiliccarrier, in which the micelles have an average diameter of less thanabout 100 nm. More preferred embodiments provide micelles having anaverage diameter less than about 50 nm, and even more preferredembodiments provide micelles having an average diameter less than about30 nm, or even less than about 20 nm.

While all suitable amphiphilic carriers are contemplated, the presentlypreferred carriers are generally those that haveGenerally-Recognized-as-Safe (GRAS) status, and that can both solubilizethe compound of the present invention and microemulsify it at a laterstage when the solution comes into a contact with a complex water phase(such as one found in human gastro-intestinal tract). Usually,amphiphilic ingredients that satisfy these requirements have HLB(hydrophilic to lipophilic balance) values of 2-20, and their structurescontain straight chain aliphatic radicals in the range of C-6 to C-20.Examples are polyethylene-glycolized fatty glycerides and polyethyleneglycols.

Particularly preferred amphiphilic carriers are saturated andmonounsaturated polyethyleneglycolyzed fatty acid glycerides, such asthose obtained from fully or partially hydrogenated various vegetableoils. Such oils may advantageously consist of tri-. di- and mono-fattyacid glycerides and di- and mono-polyethyleneglycol esters of thecorresponding fatty acids, with a particularly preferred fatty acidcomposition including capric acid 4-10, capric acid 3-9, lauric acid40-50, myristic acid 14-24, palmitic acid 4-14 and stearic acid 5-15%.Another useful class of amphiphilic carriers includes partiallyesterified sorbitan and/or sorbitol, with saturated or mono-unsaturatedfatty acids (SPAN-series) or corresponding ethoxylated analogs(TWEEN-series).

Commercially available amphiphilic carriers are particularlycontemplated, including Gelucire-series, Labrafil, Labrasol, orLauroglycol (all manufactured and distributed by Gattefosse Corporation,Saint Priest, France), PEG-mono-oleate, PEG-di-oleate, PEG-mono-laurateand di-laurate, Lecithin, Polysorbate 80, etc (produced and distributedby a number of companies in USA and worldwide).

Hydrophilic polymers suitable for use in the present invention are thosewhich are readily water-soluble, can be covalently attached to avesicle-forming lipid, and which are tolerated in vivo without toxiceffects (i.e., are biocompatible). Suitable polymers includepolyethylene glycol (PEG), polylactic (also termed polylactide),polyglycolic acid (also termed polyglycolide), a polylactic-polyglycolicacid copolymer, and polyvinyl alcohol. Preferred polymers are thosehaving a molecular weight of from about 100 or 120 daltons up to about5,000 or 10,000 daltons, and more preferably from about 300 daltons toabout 5,000 daltons. In a particularly preferred embodiment, the polymeris polyethyleneglycol having a molecular weight of from about 100 toabout 5,000 daltons, and more preferably having a molecular weight offrom about 300 to about 5,000 daltons. In a particularly preferredembodiment, the polymer is polyethyleneglycol of 750 daltons (PEG(750)).The polymers used in the present invention have a significantly smallermolecular weight, approximately 100 daltons, compared to the large MW of5000 daltons or greater that used in standard pegylation techniques.Polymers may also be defined by the number of monomers therein; apreferred embodiment of the present invention utilizes polymers of atleast about three monomers, such PEG polymers consisting of threemonomers (approximately 150 daltons).

Other hydrophilic polymers which may be suitable for use in the presentinvention include polyvinylpyrrolidone, polymethoxazoline,polyethyloxazoline, polyhydroxypropyl methacrylamide,polymethacrylamide, polydimethylacrylamide, and derivatized cellulosessuch as hydroxymethylcellulose or hydroxyethylcellulose.

In certain embodiments, a formulation of the present invention comprisesa biocompatible polymer selected from the group consisting ofpolyamides, polycarbonates, polyalkylenes, polymers of acrylic andmethacrylic esters, polyvinyl polymers, polyglycolides, polysiloxanes,polyurethanes and co-polymers thereof, celluloses, polypropylene,polyethylenes, polystyrene, polymers of lactic acid and glycolic acid,polyanhydrides, poly(ortho)esters, poly(butic acid), poly(valeric acid),poly(lactide-co-caprolactone), polysaccharides, proteins, polyhyaluronicacids, polycyanoacrylates, and blends, mixtures, or copolymers thereof.

The release characteristics of a formulation of the present inventiondepend on the encapsulating material, the concentration of encapsulateddrug, and the presence of release modifiers. For example, release can bemanipulated to be pH dependent, for example, using a pH sensitivecoating that releases only at a low pH, as in the stomach, or a higherpH, as in the intestine. An enteric coating can be used to preventrelease from occurring until after passage through the stomach. Multiplecoatings or mixtures of cyanamide encapsulated in different materialscan be used to obtain an initial release in the stomach, followed bylater release in the intestine. Release can also be manipulated byinclusion of salts or pore forming agents, which can increase wateruptake or release of drug by diffusion from the capsule. Excipientswhich modify the solubility of the drug can also be used to control therelease rate. Agents which enhance degradation of the matrix or releasefrom the matrix can also be incorporated. They can be added to the drug,added as a separate phase (i.e., as particulates), or can beco-dissolved in the polymer phase depending on the compound. In allcases the amount should be between 0.1 and thirty percent (w/w polymer).Types of degradation enhancers include inorganic salts such as ammoniumsulfate and ammonium chloride, organic acids such as citric acid,benzoic acid, and ascorbic acid, inorganic bases such as sodiumcarbonate, potassium carbonate, calcium carbonate, zinc carbonate, andzinc hydroxide, and organic bases such as protamine sulfate, spermine,choline, ethanolamine, diethanolamine, and triethanolamine andsurfactants such as Tween®. and Pluronic®. Pore forming agents which addmicrostructure to the matrices (i.e., water soluble compounds such asinorganic salts and sugars) are added as particulates. The range shouldbe between one and thirty percent (w/w polymer).

Uptake can also be manipulated by altering residence time of theparticles in the gut. This can be achieved, for example, by coating theparticle with, or selecting as the encapsulating material, a mucosaladhesive polymer. Examples include most polymers with free carboxylgroups, such as chitosan, celluloses, and especially polyacrylates (asused herein, polyacrylates refers to polymers including acrylate groupsand modified acrylate groups such as cyanoacrylates and methacrylates).

The above-described administration schedules are provided forillustrative purposes only and should not be considered limiting. Aperson of ordinary skill in the art will readily understand that alldoses are within the scope of the invention.

Compounds of the invention can be advantageously administered withsecond agents to patients in need thereof. When a rho-kinase inhibitoris administered with a second agent, the rho-kinase inhibitor and thesecond agent can be adminstered sequentially or concomitantly.Sequentially means that one agent is administered for a time followed byadministration of the second agent, which may be followed byadministration of the first agent. When agents are administeredsequentially, the level or one agent may not be maintained at atherapeutically effective level when the second agent is administered,and vice versa. Concomitantly means that the first and second agent areadministered according to a schedule that maintains both agents at ansubstantially therapeutically effective level, even though the agentsare not administered simultaneously. Each agent can be administered insingle or multiple doses, and the doses can be administered on anyschedule, including, without limitation, twice daily, daily, weekly,every two weeks, and monthly.

The invention also includes adjunctive administration. Adjunctiveadministration means that a second agent is administered to a patient inaddition to a first agent that is already being administered to treat adisease or disease symptom. In some embodiments, adjunctiveadministration involves administering a second agent to a patient inwhich administration of the first agent did not sufficiently treat adisease or disease symptom. In other embodiments, adjunctiveadministration involves administration of the second agent to a patientwhose disease has been effectively treated by administration of thefirst agent, with the expectation that the adjunctive treatment improvesthe outcome of the treatment. In some embodiments, the effect ofadministering the first and second agents is synergistic. In someembodiments, administration of the first and second agents prevents orlengthens the time until relapse, compared to administration of eitherof the agents alone. In some embodiments, administration of the firstand second agents allows for reduced dosage and/or frequency ofadministration of the first and second agent.

In an embodiment of the invention, a rho-kinase inhibitor of theinvention is administered and an anti-neoplastic agent are administeredto a subject in need thereof. In another embodiment, a rho-kinaseinhibitor of the invention and an angiogenesis inhibitor areadministered to a subject in need thereof. In another embodiment, arho-kinase inhibitor of the invention and an anti-inflammatory agent areadministered to a subject in need thereof. In yet another embodiment, arho-kinase inhibitor of the invention and an immunosuppressant areadministered. The second agent can be, without limitation, a smallmolecule, an antibody or antigen binding fragment thereof, or radiation.

Antineoplastic agents include, without limitation, cytotoxicchemotherapeutic agents, targeted small molecules and biologicalmolecules, and radiation. Compounds and agents that can be administeredfor oncological treatment, in addition to a rho kinase inhibitor of theinvention, include the following: irinotecan, etoposide, camptothecin,5-fluorouracil, hydroxyurea, tamoxifen, paclitaxel, capcitabine,carboplatin, cisplatin, bleomycin, dactomycin, gemcitabine, doxorubicin,danorubicin, cyclophosphamide, and radiotherapy, which can be external(e.g., external beam radiation therapy (EBRT)) or internal (e.g.,brachytherapy (BT)).

Targeted small molecules and biological molecules include, withoutlimitation, inhibitors of components of signal transduction pathways,such as modulators of tyrosine kinases and inhibitors of receptortyrosine kinases, and agents that bind to tumor-specific antigens.Examples include inhibitors of epidermal growth factor receptor (EGFR),including gefitinib, erlotinib, and cetuximab, inhibitors of HER2 (e.g.,trastuzumab, trastuzumab emtansine (trastuzumab-DM1; T-DM1) andpertuzumab), anti-VEGF antibodies and fragments (e.g., bevacizumab),antibodies that inhibit CD20 (e.g., rituximab, ibritumomab), anti-VEGFRantibodies (e.g., ramucirumab (IMC-1121B), IMC-1C11, and CDP791),anti-PDGFR antibodies, and imatinib. Small molecule kinase inhibitorscan be specific for a particular tyrosine kinase or be inhibitors of twoor more kinases. For example, the compoundN-(3,4-dichloro-2-fluorophenyl)-7-({[(3aR,6aS)-2-methyloctahydrocyclopenta[c]pyrrol-5-yl]methyl}oxy)-6-(methyloxy)quinazolin-4-amine(also known as XL647, EXEL-7647 and KD-019) is an in vitro inhibitor ofseveral receptor tyrosine kinases (RTKs), including EGFR, EphB4, KDR(VEGFR), Flt4 (VEGFR3) and ErbB2, and is also an inhibitor of the SRCkinase, which is involved in pathways that result in nonresponsivenessof tumors to certain TKIs. In an embodiment of the invention, treatmentof a subject in need comprises administration of a rho-kinase inhibitorof Formula I and administration of KD-019.

Dasatinib (BMS-354825; Bristol-Myers Squibb, New York) is another orallybioavailable, ATP-site competitive Src inhibitor. Dasatanib also targetsBcr-Abl (FDA-approved for use in patients with chronic myelogenousleukemia (CML) or Philadelphia chromosome positive (Ph+) acutelymphoblastic leukemia (ALL)) as well as c-Kit, PDGFR, c-FMS, EphA2, andSrc family kinases. Two other oral tyrosine kinase inhibitor of Src andBcr-Abl are bosutinib (SKI-606) and saracatinib (AZD0530).

According to the invention, angiogenesis inhibitors can be administeredto a subject in conjunction with compounds of the invention.Angiogenesis inhibitors include any substance that inhibits the growthof new blood vessels. For example, angiogenesis inhibitors includeantagonists of VEGF, PlGF, and VEGF receptors, including the antibodiesdisclosed herein. By inhibitor is meant an inhibitor of a biologicalprocess or inhibitor of a target. In this regard, an angiogenesisinhibitor is an agent that reduces angiogenesis. A Rho-kinase inhibitoris an agent, such as a competitive inhibitor of ATP binding, thatinhibits an intrinsic activity or blocks an interaction of Rho-kinase.By antagonist is meant a substance that reduces or inhibits an activityor function in a cell associated with a target. For example, a VEGFantagonist reduces or blocks a function in a cell that is associatedwith VEGF. A VEGF antagonist may act on VEGF, by binding to VEGF andblocking binding to its receptors and/or may act on another cellularcomponent involved in VEGF-mediated signal transduction. Similarly, aVEGFR2 antagonist is an agent that reduces or blocks VEGFR2-mediatedsignal transduction by binding to VEGFR2 and blocking ligand binding orinteraction with a VEGFR2 substrate, or acts on another cellularcomponent to reduce or block VEGFR2-mediated signal transduction. Thus,angiogenesis inhibitors include antagonists of, without limitation,VEGF, VEGFR1, VEGFR2, PDGF, PDGFR-β, neuropilin-1 (NRP1), andcomplement.

Non-limiting examples of VEGF-binding agents include VEGF antibodies andVEGF traps (i.e., ligand binding domains of VEGF receptors. In general,a VEGF trap is a protein that comprises VEGF binding domains of one ormore VEGF receptor protein. VEGF-traps include, without limitation,soluble VEGFR-1, soluble neuropilin 1 (NRP1), soluble VEGFR-3 (whichbinds VEGF-C and VEGF-D), and aflibercept (Zaltrap; Eyelea; VEGF TrapR1R2), comprised of segments of the extracellular domains of humanvascular endothelial growth factor receptors VEGFR1 and VEGFR2 fused tothe constant region (Fc) of human IgG1. Conbercept (KH902) is a fusionprotein which contains the extracellular domain 2 of VEGFR-1 (Flt-1) andextracellular domain 3, 4 of VEGFR-2 (KDR) fused to the Fc portion ofhuman IgG1. Several VEGF traps containing KDR and FLT-1 Ig-like domainsin various combinations are disclosed in U.S. Pat. No. 8,216,575.DARPins (an acronym for designed ankyrin repeat proteins) aregenetically engineered antibody mimetic proteins typically exhibitinghighly specific and high-affinity target protein binding. DARPin® MP0112is a vascular endothelial growth factor (VEGF) inhibitor and has enteredclinical trials for the treatment of wet macular degeneration anddiabetic macular edema.

According to the invention, VEGF expression can be targeted. Forexample, VEGF inhibitor PTC299 targets VEGF post-transcriptionally byselectively binding the 5′- and 3′-untranslated regions (UTR) of VEGFmessenger RNA (mRNA), thereby preventing translation of VEGF. Pegaptanib(Macugen) is an RNA aptamer directed against VEGF-165.

Placental growth factor (PlGF) has been implicated in pathologicalangiogenesis. PlGF is structurally related to VEGF and is also a ligandfor VEGFR-1. Consequently, VEGF traps comprising the extracellulardomain of VEGFR1 (see above) are useful for targeting PlGF.

PDGF is composed of four polypeptide chains that form homodimersPDGF-AA, BB, CC, and DD as well as the heterodimer PDGF-AB. The PDGFreceptors (PDGFR)-α and -β mediate PDGF functions. Specifically, PDGFRαbinds to PDGF-AA, -BB, -AB, and -CC, whereas PDGFRβ interacts with -BBand -DD. Non-limiting examples of PDGF-binding agents include anti-PDGFantibodies and PDGF traps. Agents that target PDGF include Fovista™(E10030, Ophthotech), a pegylated aptamer targeting PDGF-B, and AX102(Sennino et al., 2007, Cancer Res. 75(15):7359-67), a DNAoligonucleotide aptamer that binds PDGF-B.

Agents that target PDGF receptors include ramucirumab (IMC-3G3, humanIgG₁) an anti-PDGFRα antibody, crenolanib (CP-868596), a selectiveinhibitor of PDGFRα(IC₅₀=0.9 nM) and PDGFRβ (IC₅₀=1.8 nM), and nilotinib(Tasigna®), an inhibitor of PDGFRα and PDGFRβ and other tyrosinekinases.

Angiogenesis inhibitors include intracellular agents that block signaltransduction mediated by, for example, VEGF, PDGF, ligands of VEGF orPDGF receptors, or complement. Intracellular agents that inhibitangiogenesis inhibitors include the following, without limitation.Sunitinib (Sutent; SU11248) is a panspecific small-molecule inhibitor ofVEGFR1-VEGFR3, PDGFRα and PDGFRβ, stem cell factor receptor (cKIT),Flt-3, and colony-stimulating factor-1 receptor (CSF-1R). Axitinib(AG013736; Inlyta) is another small molecule tyrosine kinase inhibitorthat inhibits VEGFR-1-VEGFR-3, PDGFR, and cKIT. Cediranib (AZD2171) isan inhibitor of VEGFR-1-VEGFR-3, PDGFRβ, and cKIT. Sorafenib (Nexavar)is another small molecular inhibitor of several tyrosine proteinkinases, including VEGFR, PDGFR, and Raf kinases. Pazopanib (Votrient;(GW786034) inhibits VEGFR-1, -2 and -3, cKIT and PDGFR. Foretinib(GSK1363089; XL880) inhibits VEGFR2 and MET. CP-547632 is as a potentinhibitor of the VEGFR-2 and basic fibroblast growth factor (FGF)kinases. E-3810 ((6-(7-((1-aminocyclopropyl)methoxy)-6-methoxyquinolin-4-yloxy)-N-methyl-1-naphthamide) inhibitsVEGFR-1, -2, and -3 and FGFR-1 and -2 kinases in the nanomolar range.Brivanib (BMS-582664) is a VEGFR-2 inhibitor that also inhibits FGFreceptor signaling. CT-322 (Adnectin) is a small protein based on ahuman fibronectin domain and binds to and inhibits activation of VEGFR2.Vandetanib (Caprelas; Zactima; ZD6474) is an inhibitor of VEGFR2, EGFR,and RET tyrosine kinases. X-82 (Xcovery) is a small molecule indolinoneinhibitor of signaling through the growth factor receptors VEGFR andPDGFR

Anti-inflammatories and immunosuppressants include steroid drugs such asglucocorticoids (e.g., dexamethasone), FK506 (tacrolimus), ciclosporin,fingolimod, interferon, such as IFNβ or IFNγ, a tumor necrosisfactor-alpha (TNF-α) binding protein such as infliximab (Remicade),etanercept (Enbrel), or adalimumab (Humira), and mycophenolic acid.

In certain embodiments, ROCK inhibitors of the invention arecoadministered with agents used to treat metabolic disorders. Forexample, for treatment of obesity, the ROCK inhibitors may be combinedwith weight loss drugs such as, but not limited to, phentermine, fatadsorption inhibitors (e.g., Xenical), appetite suppressants, and thelike. Procedures used to assist weight loss include, for example,stomach bands, stomach bypass or stapling. For insulin resistance ormetabolic syndrome or hyperinsulinemia, ROCK inhibitors of the inventioncan be coadministered with compounds that lower cholesterol levels, forexample, one or more medicines such as statins, fibrates, or nicotinicacid. For high blood pressure associated with such diseases, ROCKinhibitors of the invention can be coadministered with, for example, oneor more antihypertensive medicines such as diuretics orangiotensin-converting enzyme (ACE) inhibitors. ROCK inhibitors of theinvention can be administered in a treatment program that includeslifestyle changes such as increased physical activity, an improved diet,and/or quitting smoking. In certain embodiments, the ROCK inhibitor isROCK2 selective.

Th17 cells are novel subset of helper CD4⁺ T cells that secrete IL-17,IL-21 and IL-22. The pro-inflammatory activity of Th17 cells can bebeneficial to the host during infection, but uncontrolled Th17 functionhas been linked and actively involved in several autoimmune pathologies.Indeed, high levels of IL-17 are detected in the sera and biopsies ofrheumatoid arthritis (RA) and systemic lupus erythematosus (SLE)patients which correlates with destruction of synovial tissue anddisease activity. The pathological role of IL-17 in arthritic joints isassociated with its stimulation of pro-inflammatory cytokine productionand increased recruitment of T cells and innate immune cells. Moreover,numbers of Th17 cells are significantly increased in the peripheralblood of RA patients as well as elevated concentrations of IL-17 wereseen in supernatants of their PBMCs after stimulation with anti-CD3/CD28antibodies ex vivo. In addition, in multiple sclerosis (MS) patients,myelin reactive Th17 cells are also enriched and produce high amounts ofIL-22 and IFN-γ. Further, a significantly higher number of IL-17⁺ cellsis detected in disease-affected gut areas compared to healthy areas ofthe same subjects with Crohn's disease (CD).

The development and function of Th17 cells depends on activation ofspecific intracellular signaling pathways. The steroid receptor-typenuclear receptor RORγt is selectively expressed in Th17 cells andappears to be required for IL-17 production. The induction of RORγt hasbeen observed to be mediated by IL-6, IL-21 and IL-23 via aSTAT3-dependent mechanism. STAT3 also binds directly to the IL-17 andIL-21 promoters. In addition to RORγt and STAT3, the interferonregulatory factor 4 (IRF4) is required for the differentiation of Th17cells since IRF4 KO mice failed to mount Th17 response and wereresistant to development of autoimmune responses. Recent studies havedemonstrated that phosphorylation of IRF4 by Rho-kinase 2 (ROCK2)regulates IL-17 and IL-21 production and development of autoimmunity inmice.

According to the invention, targeting Th17 (IL-17-secreting) cells byrho-kinase inhibition provides a method for treating Th17 cell-mediateddiseases, including but not limited to autoimmune disorders such asrheumatoid arthritis (RA) multiple sclerosis (MS), systemic luypussrythematosus (SLE), psoriasis, Crohn's disease, atopic dermatitis,eczema, and GVHD in humans. In an embodiment of the invention, theRho-kinase inhibitor is a compound of Formula I. In some embodiments,the rho-kinase inhibitor inhibits ROCK1 and ROCK2. In some embodiments,the rho-kinase inhibitor selectively inhibits ROCK2. Selectiveinhibition of ROCK2 provides for treatment of Th17 cell-mediateddiseases and reduces or prevents toxicities associated with completeinhibition of ROCK activity.

Regulatory T cells (Tregs) play a critical role in the maintenance ofimmunological tolerance to self-antigens and inhibition of autoimmuneresponses, but, at the same time, prevent an effective immune responseagainst tumor cells. Indeed, Tregs isolated from the peripheral blood ofpatients with autoimmune disease, such as rheumatoid arthritis (RA) andmultiple sclerosis (MS), show a defect in their ability to suppresseffector T cell function, while increased accumulation of Tregscorrelates with a poor prognosis in many cancers. Thus, the level ofTreg function effects a balance between effective immunity and avoidanceof pathological autoreactivity.

The development and function of Tregs depend on activation of specificsignaling transduction pathways. TGF-β and IL-2 activate expression ofFoxp3 and STAT5 transcription factors that both play an essential rolein the control of Treg suppressive function. On the other hand,pro-inflammatory cytokines inhibit Foxp3 expression via up-regulation ofSTAT3 phosphorylation. As shown herein, pharmacological inhibition ofROCK2 (e.g., with selective ROCK2 inhibitors such as KD025,ROCK2-specific siRNA-mediated inhibition of ROCK2), but not ROCK1, leadsto down-regulation of STAT3 phosphorylation, interferon regulatoryfactor 4 (IRF4) and steroid receptor-type nuclear receptor RORγt proteinlevels in human T cells.

Furthermore, as demonstrated herein, targeting ROCK2 with a selectiveinhibitor (e.g., KD025) leads to an increased proportion of Foxp3⁺ Tcells via a STAT5-dependent mechanism and positively regulates theirsuppressive activity towards autoreactive lymphocytes. This effect ofROCK2 inhibition on Tregs is critical to limiting or preventing theonset of aberrant self-immune responses. This effect may be shown, forexample, by assaying the ability of Tregs treated with a ROCK2 inhibitorto inhibit proliferation and cytokine secretion in target cells invitro.

Accordingly, ROCK2 inhibitors of the invention effectively reduce orprevent chronic GVHD pathologies. As shown herein, the compounds of theinvention reduce or prevent cGVHD pathologies in target organs. Forexample, a selective ROCK2 inhibitor administered to a transplantpatient maintains or restores pulmonary function. The maintenance orrestoration of pulmonary function correlates with reduced germinalcenter activity which would otherwise lead to production of autoreactiveantibodies, and with reduced collagen deposition and antibody depositionin affected tissues. These pathologies are common to other targets ofcGVHD as well, e.g., skin, gut, and liver, which would be similarlyreduced or prevented.

It is to be understood and expected that variations in the principles ofinvention herein disclosed may be made by one skilled in the art and itis intended that such modifications are to be included within the scopeof the present invention.

Throughout this application, various publications are referenced. Thesepublications are hereby incorporated into this application by referencein their entireties to more fully describe the state of the art to whichthis invention pertains. The following examples further illustrate theinvention, but should not be construed to limit the scope of theinvention in any way.

EXAMPLES

Abbreviations used in the following examples and preparations include:

Ac₂O acetic anhydride

AcOH acetic acid

Bn Benzyl

Celite® diatomaceous earth

DCM dichloromethane

DIEA di-isopropylethylamine

DMAP 4-dimethylamino pyridine

DME 1,2-dimethoxylethane

DMF dimethylformamide

DMSO dimethyl sulfoxide

EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodimide hydrochloride

EtOAc ethyl acetate

EtOH ethyl alcohol or ethanol

Et₂O ethyl ether

Et₃N triethylamine

g grams

HOBt 1-hydroxybenzotriazole

HPLC high pressure liquid chromatography

h hour(s)

MeCN acetonitrile

min minute(s)

MeOH methyl alcohol or methanol

mL milliliter

mmol millimoles

MS mass spectrometry

NMR nuclear magnetic resonance

iPrOH iso-propanol

PyBOP® benzotriazol-1-yl-oxytripyrrolidinophosphonium

rt room temperature

s singlet

t triplet

THF tetrahydrofuran

Mass spectrometry was conducted by: SynPep Co., 6905 Ct. Dublin, Calif.94568, or it was recorded on an LC-MS: Waters 2695 Separations Modulewith a Waters ZQ2000 single quadrapole MS detector. Unless stated allmass spectrometry was run in ESI mode. ¹H NMR spectra were recorded on aVarian 400 MHz machine using Mercury software. In sofar the synthesis ofthe following examples of compounds of the present invention is notexplicitly described in such example, the synthesis is as describedherein in general terms and the appropriate starting material can beeasily selected for synthesizing the compound of the example.

Example 1 2-Bromo-N-isoopropylacetamide

A solution of iso-propyl amine (5.0 g, 7.20 mL, 84.6 mmol) in 63 mL ofDCM was cooled to −10° C. To this was added a solution ofα-bromoacetylbromide (8.53 g, 3.68 mL, 42.3 mmol) in 10.5 mL of DCM. Thereaction mixture was stirred for 10 min. The iso-propylammoniumhydrobromide was filtered from the mixture and the filtrate thenconcentrated in vacuo to give the title compound as a white solid (5.30g, 70%).

Example 2N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide

A mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (0.50g, 2.27 mmol), 2-bromo-N-isoopropylacetamide (0.61 g, 3.41 mmol), andK₂CO₃ (0.47 g, 3.41 mmol) in DMF (3 mL) was stirred at rt followed byaddition of ice water. The precipitate was filtered and washed withwater and dried to provide the title compound (0.32 g, 44%).

Example 3 tert-Butyl5-((2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of 2,4-dichloropyrimidine (1.99 g, 13.4 mmol), tert-butyl5-amino-1H-indazole-1-carboxylate (3.4 g, 14.7 mmol), DIEA (3 mL), andDMF (13 mL) was stirred at 65° C. for 7 h, concentrated in vacuo, andtiturated with Et₂O. The precipitate was filtered and washed with IPAand dried to provide the title compound (1.83 g, 40%).

Example 4 tert-Butyl5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate (100 mg,0.29 mmol),N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(130 mg, 0.41 mmol), Pd(dppf)Cl₂.CH₂Cl₂ (20 mg, 0.02 mmol), and K₂CO₃(80 mg, 0.58 mmol) in dioxane/water (10 and 2 mL) was heated inmicrowave for 30 min. The reaction was worked up and purified bychromatography to provide the title compound (176 mg, 35%).

Example 52-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideHCl salt

tert-Butyl5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylatewas taken up in 4 M HCl in dioxane and stirred at rt for 2 h. Thevolatiles were removed in vacuo to give the title compound as HCl salt.¹H NMR (400 MHz, DMSO-d₆) δ12.6 (d, J=6.8 Hz, 6H), 3.93 (m, 1H), 4.51(s, 2H), 6.75 (d, J=6 Hz, 1H), 7.14 (d, J=7.6 Hz, 1H), 7.44-7.59 (m,3H), 7.87-7.91 (m, 3H), 8.09 (s, 1H), 8.18 (s, 1H), 8.31 (d, J=6.4 Hz,1H), 10.19 (d, J=0.8 Hz, 1H), 13.10 (s, 1H). MS (ES+) m/e 403 (M+H)⁺.

Example 6 tert-butyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetate

3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol, tert-butyl2-bromoacetate

A mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (4 g,18.2 mmol), tert-butyl 2-bromoacetate (5.7 g, 27.3 mmol) and K₂CO₃ (3.44g, 27.3 mmol) in CH₃CN (100 mL) was stirred at 70° C. overnight. Thereaction mixture was poured into water and extracted with EtOAc. Theorganic layer was dried over Na₂SO₄ and removed to give a residue. Theresidue was purified by column chromatograph to give the title compound(4 g, 67%) as a white solid.

Example 7 N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine

A mixture of compound 2,4-dichloropyrimidine (14.8 g, 0.1 mol),1H-indazol-5-amine (14.6 g, 110 mmol) and Et₃N (15 g, 150 mmol) in EtOH(200 mL) was stirred at 80° C. for 3 hrs. The reaction mixture wascooled and filtered. The filtered cake was collected and dried to givecompound 5 (15 g, 60%) as a solid.

Example 8 tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (7.35 g,30 mmol), Boc₂O (18.9 g, 90 mmol) in CH₂Cl₂ (150 mL) was added DMAP (3.6g, 30 mmol) at 0° C. during 5 min. After 0.5 hr, the reaction wascompleted. The reaction mixture was washed with water, dried over Na₂SO₄and removed to give a residue, which was purified by gel columnchromatograph to give the title compound (6 g, 67%) as a white solid.

Example 9 tert-butyl5-((2-(3-(2-(tert-butoxy)-2-oxoethoxy)phenyl)pyrimidin-4-yl)(tert-butoxycarbonyl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(4 g, 8.9 mmol), tert-butyl2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetate (3.3g, 10 mmol), KOAc (35 g, 360 mmol), Pd(dppf)₂Cl₂ (400 mg) and Boc₂O (3.9g, 18 mmol) in dioxane/water (10/1, 100 mL) was stirred at 100° C. for 3days. The reaction mixture was poured into water and extracted withEtOAc. The organic layer was dried over Na₂SO₄ and removed to give aresidue, which was purified by gel column chromatograph to give thetitle compound (3 g, 54%) as a solid.

Example 10 2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)aceticacid

A mixture of compound tert-butyl5-((2-(3-(2-(tert-butoxy)-2-oxoethoxy)phenyl)pyrimidin-4-yl)(tert-butoxycarbonyl)amino)-1H-indazole-1-carboxylate(2 g) and CF₃COOH (20 mL) in DCM (20 mL) was stirred at 25° C. for 2hrs. The solvent was removed to give the title compound (1.5 g) as ayellow solid.

Example 11 (S)-tert-butyl3-(2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetamido)pyrrolidine-1-carboxylate

A mixture of2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetic acid (600mg, 1.66 mmol), 3-amino-pyrrolidine-1-carboxylic acid tert-butyl ester(300 mg, 1.62 mmol), HATU (760 mg, 2 mmol) and Et₃N (250 mg, 2 mmol) inDMF (18 mL) was stirred at 25° C. overnight. The reaction mixture waspoured into water and extracted with EtOAc. The organic layer was driedover Na₂SO₄ and concentrated to give a residue, which was purified byHPLC to provide the title compound (300 mg, 50%) as a solid.

Example 12(R)-2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-N-(pyrrolidin-3-yl)acetamideHCl salt

¹H NMR (400 MHz, DMSO-d₆) δ1.84-1.96 (m, 1H), 2.07-2.19 (m, 1H),3.07-3.38 (m, 4H), 4.40-4.42 (m, 1H), 4.65 (s, 2H), 7.36 (d, J=6.4 Hz,1H), 7.56 (t, J=8.0 Hz, 1H), 7.65 (d, J=8.0 Hz, 1H), 7.90-7.94 (m, 2H),8.14-8.29 (m, 2H), 8.66 (d, J=6.8 Hz, 1H), 9.39-9.80 (m, 3H), 11.87 (s,1H). MS (ES+) m/e 430 (M+H)⁺.

Example 13(R)-2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-N-(pyrrolidin-3-yl)acetamideHCl salt

A mixture of compound2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetic acid (500mg, 1.39 mmol), NH₄Cl (125 mg, 2 mmol), HATU (720 mg, 1.89 mmol) andEt₃N (200 mg, 2 mmol) in DMF (15 mL) was stirred at 25° C. overnight.The reaction mixture was poured into water and extracted with EtOAc. Theorganic layer was dried over Na₂SO₄ and concentrated to give a residue,which was purified by HPLC to provide the title compound (60.1 mg, 12%)as a solid. ¹H NMR (400 MHz, DMSO-d₆) δ4.52 (s, 2H), 6.78 (d, J=6.4 Hz,1H), 7.19 (dd, J=9.6 and 1.6 Hz, 1H), 7.40-7.62 (m, 5H), 7.84-7.87 (m,2H), 8.11 (s, 1H), 8.15 (s, 1H), 8.33 (d, J=6.4 Hz, 1H), 10.35 (s, 1H).MS (ES+) m/e 361 (M+H)⁺.

Example 142-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-1-(1,1-dioxidothiomorpholino)ethanoneTFA salt

A mixture of compound2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetic acid (500mg, 1.39 mmol), thiomorpholine 1,1-dioxide (375 mg, 2.2 mmol), HATU (720mg, 1.89 mmol) and Et3N (200 mg, 2 mmol) in DMF (15 mL) was stirred at25° C. overnight. The reaction mixture was poured into water andextracted with EtOAc. The organic layer was dried over Na2SO4 andconcentrated to give a residue, which was purified by pre-HPLC to givethe title compound (54.6 mg, 10%) as a solid. 1H NMR (400 MHz, DMSO-d6)δ3.11 (b, 2H), 3.30 (b, 2H), 3.85 (b, 4H), 5.00 (s, 2H), 6.76 (d, J=6.4Hz, 1H), 6.91 (s, 1H), 7.09 (s, 1H), 7.16 (dd, J=8.0 and 2.4 Hz, 1H),7.45 (t, J=8.0 Hz, 1H), 7.47-7.50 (m, 2H), 7.81 (s, 1H), 7.86 (d, J=8.0Hz, 1H), 8.08 (s, 1H), 8.12 (b, 1H), 8.33 (d, J=6.4 Hz, 1H), 10.26 (s,1H). MS (ES+) m/e 479 (M+H)⁺.

Example 15N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine-4-carboxamide

To a mixture of triphosgene (600 mg, 2 mmol) in THF (10 mL) was droppeda solution of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline(836 mg, 4 mmol) and Et₃N (1.2 g, 12 mmol) in THF (10 mL) during 10 minat 40° C. After 0.5 hr, morpholine (435 mg, 5 mmol) was added to thereaction mixture. After 15 min, the reaction mixture was quenched withsaturated NH₄Cl, extracted with EtOAc. The organic layer was dried overNa₂SO₄ and concentrated to give a residue, which was purified by gelcolumn chromatography to give the title compound (530 mg, 50%) as awhite solid.

Example 16 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(morpholine-4-carboxamido)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(300 mg, 0.7 mmol),N-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)morpholine-4-carboxamide(250 mg, 0.8 mmol), CsF (510 mg, 3 mmol), Pd(PPh₃)₄ (120 mg) and Boc₂O(432 mg, 2 mmol) in dioxane/water (10/1, 10 mL) was stirred at 130° C.under microwave for 20 min. Three pots were combined. The reactionmixture was diluted with water and extracted with EtOAc. The organiclayer was dried over Na₂SO₄ and concentrated to give a residue, whichwas purified by column chromatograph to give the title compound (360mg).

Example 17N-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenyl)morpholine-4-carboxamideHCl salt

A mixture of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(morpholine-4-carboxamido)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(360 mg, crude) in MeOH/HCl (4M, 20 mL) was stirred at 25° C. overnight.The reaction mixture was concentrated to give title compound (68 mg) asan HCl salt. ¹H NMR (400 MHz, DMSO-d₆) δ3.49 (s, 4H), 3.62 (s, 4H), 7.14(b, 1H), 7.50 (t, J=7.6 Hz, 1H), 7.60-7.65 (m, 3H), 7.88 (d, J=6.8 Hz,1H), 8.22 (s, 1H), 8.28 (d, J=5.6 Hz, 1H), 8.46 (b, 1H), 8.68 (s, 1H),9.00 (s, 1H), 11.77 (s, 1H). MS (ES+) m/e 416 (M+H)⁺.

Example 18 Methyl 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzoate

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (3.7 g, 15mmol), 3-methoxycarbonylphenylboronic acid (3.3 g, 18 mmol), K₂CO₃ (4.14g, 30 mmol) and Pd(dppf)₂Cl₂ (700 mg) in dioxane/water (4/1, 75 mL) wasstirred at 100° C. for 16 hrs. The reaction mixture was concentrated togive the title compound (7 g, crude) which was carried out directly fornext step reaction without further purification.

Example 19 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzoic acid

To a mixture of methyl3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzoate (7 g, crude) indioxane (120 mL) was added NaOH (2M, 120 mL). The reaction mixture wasrefluxed for 1.5 h and was then cooled to rt and extracted with EtOAc(100 mL). The aqueous phase was separated and acidified with HCl (6M).The mixture was filtered and the filter cake was collected, dried togive the title compound (1.3 g, crude) used for the next step reactiondirectly.

Example 203-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)-N-(1,1-dioxidotetrahydro-2H-thiopyran-4-yl)benzamideTFA salt

To the mixture of 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzoicacid (400 mg, 1.2 mmol), 4-aminotetrahydro-2H-thiopyran 1,1-dioxide (300mg, 1.4 mmol), HATU (720 mg, 1.89 mmol) and Et₃N (200 mg, 2 mmol) in DMF(15 mL) was stirred at 25° C. overnight. The reaction mixture was pouredinto water and extracted with EtOAc. The organic layer was dried overNa₂SO₄ and concentrated to give a residue, which was purified by HPLC togive the title compound (81 mg, 20%) as a solid. ¹H NMR (400 MHz, CD₃OD)δ2.19-2.24 (m, 2H), 2.31-2.33 (m, 2H), 3.12-3.15 (m, 2H), 3.34-3.38 (m,2H), 4.23-4.28 (m, 1H), 6.93 (d, J=6.8 Hz, 1H), 7.58-7.76 (m, 3H), 8.12(s, 1H), 8.14 (s, 1H), 8.24 (d, J=7.2 Hz, 1H), 8.31 (d, J=8.0 Hz, 1H),8.65 (s, 1H). MS (ES+) m/e 463 (M+H)⁺.

Example 21 tert-Butyl4-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzamido)piperidine-1-carboxylate

A mixture of 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzoic acid(400 mg, 1.2 mmol), 4-amino-piperidine-1-carboxylic acid tert-butylester (300 mg, 1.5 mmol), HATU (720 mg, 1.89 mmol) and Et₃N (200 mg, 2mmol) in DMF (15 mL) was stirred at 25° C. overnight. The reactionmixture was poured into water and extracted with EtOAc. The organiclayer was dried over Na₂SO₄ and concentrated to give a residue, whichwas purified by HPLC to provide the title compound (200 mg, 30%) as asolid.

Example 223-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)-N-(piperidin-4-yl)benzamideHCl salt

A mixture of tert-butyl4-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)benzamido)piperidine-1-carboxylate(200 mg) in HCl in MeOH (5 mL) was stirred at 25° C. for 3 h. Thesolvent was removed to provide the title compound (150 mg) as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ1.79-1.87 (m, 2H), 1.98-2.01 (m, 2H),3.01-3.04 (m, 2H), 3.32-3.36 (m, 2H), 3.64-3.69 (m, 1H), 7.10-7.15 (m,1H), 7.66 (d, J=8.0 Hz, 1H), 7.75 (t, J=8 Hz, 1H), 8.16 (s, 1H), 8.20(s, 1H), 8.36 (d, J=6.8 Hz, 1H), 8.42 (d, J=7.6 Hz, 1H), 8.78 (d, J=7.2Hz, 1H), 8.90 (s, 1H), 8.96 (s, 1H), 11.73 (s, 1H). MS (ES+) m/e 414(M+H)⁺.

Example 23 N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine

To a mixture of compound 2,4-dichloro-5-methylpyrimidine (5 g, 30.8mmol) and 5-aminoindazole (4.1 g, 30.8 mmol) in anhydrous ethanol (100mL) was added Na₂CO₃ (16 g, 154 mmol). The resulting mixture was heatedat 80° C. overnight under N₂. After cooling to room temperature, themixture was diluted with water and extracted with EtOAc. The organicphase was dried over anhydrous Na₂SO₄ and concentrated in vacuo followedby column chromatography on silica gel (eluted with DCM:MeOH=50:1) toprovide the title compound (7 g, yield 87%) as a brown solid.

Example 24 tert-butyl5-((tert-butoxycarbonyl)(2-chloro-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a solution of N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine(5 g, 19.3 mmol) in anhydrous DCM (100 mL) was added Boc₂O (12.6 g, 57.9mmol), TEA (5.85 g, 57.9 mmol) and DMAP (1.17 g, 9.56 mmol). Theresulting mixture was stirred at room temperature for 4 h. The mixturewas diluted with water and extracted with DCM. The organic phase wasdried over anhydrous Na₂SO₄, and the solvent was removed in vacuofollowed by purification by column chromatograph on silica gel (elutedwith PE:EA=10:1) to give compound the title compound (5 g, yield 56%) asa white solid.

Example 25 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(cyclopropylamino)-2-oxoethoxy)phenyl)-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a mixture of compound tert-butyl5-((tert-butoxycarbonyl)(2-chloro-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(1.9 g, 4.14 mmol), compoundN-cyclopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(1.57 g, 4.97 mmol) and CsF (1.89 g, 12.42 mmol) in 1,4-dioxane (20 mL)and water (5 mL) was added Pd(PPh₃)₄ (239 mg, 0.21 mmol). The resultingmixture was heated at 100° C. overnight under N₂. After cooling to roomtemperature, the mixture was diluted with water and extracted withEtOAc, the organic phase was dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to give a residue, which waspurified by column chromatography on silica gel (eluted with PE:EA=5:1)to provide the title compound (1.6 g, yield 62%) as a yellow solid.

Example 262-(3-(4-((1H-indazol-5-yl)amino)-5-methylpyrimidin-2-yl)phenoxy)-N-cyclopropylacetamidehydrochloride

To a solution of compound tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(cyclopropylamino)-2-oxoethoxy)phenyl)-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(500 mg, 0.81 mmol) in EtOAc (2 mL) was added HCl/EtOAc (10 mL) andstirred at room temperature overnight. The formed precipitated wasfiltered and washed with EtOAc, dried in vacuo to afford the titlecompound (300 mg, yield 89%) as a yellow solid. ¹H NMR (400 MHz, CD₃OD)δ0.50-0.54 (m, 2H), 0.70-0.75 (m, 2H), 2.42 (s, 3H), 2.66-2.71 (m, 1H),4.51 (s, 2H), 7.27 (dd, J=8.4 and 2.4 Hz, 1H), 7.49 (t, J=8.0 Hz, 1H),7.64-7.68 (m, 2H), 7.72 (s, 1H), 8.04 (s, 2H), 8.20 (s, 1H), 8.28 (s,1H). MS (ES+) m/e 451 (M+H)⁺.

Example 27 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-oxoethoxy)phenyl)-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloro-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(1.4 g, 3.05 mmol), tert-butyl4-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamido)piperidine-1-carboxylate(1.68 g, 3.66 mmol) and CsF (1.39 g, 9.15 mmol) in 1,4-dioxane (20 mL)and water (5 mL) was added Pd(PPh₃)₄ (176 mg, 0.15 mmol). The resultingmixture was heated at 100° C. overnight under N₂. After cooling to roomtemperature, the mixture was diluted with water and extracted withEtOAc, the organic phase was dried over anhydrous Na₂SO₄ andconcentrated under reduced pressure to give a residue, which waspurified by column chromatography on silica gel (eluted with PE:EA=1:1)to afford the title compound (1.1 g, yield 47%) as a yellow solid.

Example 282-(3-(4-((1H-indazol-5-yl)amino)-5-methylpyrimidin-2-yl)phenoxy)-N-(piperidin-4-yl)acetamidehydrochloride

To a solution of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-((1-(tert-butoxycarbonyl)piperidin-4-yl)amino)-2-oxoethoxy)phenyl)-5-methylpyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(700 mg, 0.92 mmol) in EtOAc (2 mL) was added HCl/EtOAc (10 mL) andstirred at room temperature overnight. The formed precipitated wasfiltered and washed with EtOAc, dried in vacuo to afford the titlecompound (230 mg, yield 54%) as a yellow solid. ¹H NMR (400 MHz, CD₃OD)δ1.73-1.84 (m, 2H), 2.01-2.05 (m, 2H), 2.42 (s, 3H), 3.02-3.09 (m, 2H),3.39-3.43 (m, 2H), 3.99-4.05 (m, 1H), 4.57 (s, 2H), 7.30 (dd, J=8.4 and2.4 Hz, 1H), 7.51 (t, J=8.4 Hz, 1H), 7.67-7.74 (m, 4H), 8.05 (s, 1H),8.20 (s, 1H), 8.24 (s, 1H). MS (ES+) m/e 494 (M+H)⁺.

Example 29 tert-butyl 2-(3-bromophenoxy)acetate

To a solution of 3-bromophenol (5 g, 28.9 mmol) in MeCN (100 mL) wasadded t-butyl bromoacetate (6.76 g, 34.7 mmol) and K₂CO₃ (5.98 g, 43.3mmol). The resulting mixture was heated at 80° C. overnight undernitrogen. After cooling to room temperature, the mixture was dilutedwith water and extracted with EtOAc. The organic layer was washed withbrine and dried over Na₂SO₄, filtrated and concentrated to give theresidue, which was purified by column chromatography on silica gel(PE:EA=50:1) to give the title compound (7 g, yield 84%) as a oilliquid.

Example 30 2-(3-bromophenoxy)acetyl chloride

To a solution of tert-butyl 2-(3-bromophenoxy)acetate (4.6 g, 16 mmol)in anhydrous DCM (50 mL) was added TFA (18 g, 0.16 mol) and stirred atroom temperature overnight. After TLC showed the reaction was completed,the mixture was concentrated under reduced pressure to get a crude acid.The acid was dissolved in anhydrous DCM (50 mL), oxalyl chloride (2.44g, 19.2 mmol) and DMF (0.2 mL) were added into the solution. The mixturewas stirred at room temperature for 4 h. The mixture was concentratedunder reduced pressure to provide a white solid, which was used for nextstep reaction without further purification.

Example 31 2-(3-bromophenoxy)-N-cyclopropylacetamide

To a solution of 2-(3-bromophenoxy)acetyl chloride (2.3 g, 9.24 mmol) inanhydrous DCM (30 mL) was added triethylamine (2.8 g, 27.7 mmol) andcyclopropylamine (632 mg, 11.1 mmol) at 0° C. Then the resulting mixturewas stirred at room temperature overnight. The reaction was diluted withwater and extracted with EtOAc. The organic layer was dried overanhydrous Na₂SO₄ and concentrated under reduced pressure to give aresidue, which was purified by column chromatography on silica gel(eluted with PE:EA=5:1) to give the title compound (1.95 g, yield 78%)as a white solid.

Example 32N-cyclopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide

To a mixture of 2-(3-bromophenoxy)-N-cyclopropylacetamide (1.95 g, 7.25mmol), bis(pinacolato)diboron (2.76 g, 10.87 mmol) and KOAc (2.13 g,21.7 mmol) in DMSO (20 mL) was added Pd(dppf)Cl₂-DCM (265 mg, 0.36mmol). The resulting mixture was heated at 100° C. overnight under N₂.After cooling to room temperature, the mixture was diluted with waterand extracted with EtOAc, the organic phase was dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to give a residue, whichwas purified by column chromatography on silica gel (eluted withPE:EA=10:1) to provide the title compound (1.4 g, yield 60%) as a whitesolid.

Example 33 tert-butyl4-(2-(3-bromophenoxy)acetamido)piperidine-1-carboxylate

To a solution of 2-(3-bromophenoxy)acetyl chloride (2.1 g, 8.43 mmol) inanhydrous DCM (30 mL) was added triethylamine (2.56 g, 25.3 mmol) and4-amino-1-Boc-piperidine (2.02 g, 10.1 mmol) at 0° C. Then the resultingmixture was stirred at room temperature overnight. The reaction wasdiluted with water and extracted with EtOAc. The organic layer was driedover anhydrous Na₂SO₄ and concentrated under reduced pressure to give aresidue, which was purified by column chromatography on silica gel(eluted with PE:EA=5:1) to give the tile compound (2.8 g, yield 80%) asa white solid.

Example 34 tert-butyl4-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamido)piperidine-1-carboxylate

To a mixture of tert-butyl4-(2-(3-bromophenoxy)acetamido)piperidine-1-carboxylate (2.8 g, 6.79mmol), bis(pinacolato)diboron (2.59 g, 10.2 mmol), and KOAc (1.99 g,20.4 mmol) in DMSO (30 mL) was added Pd(dppf)Cl₂-DCM (249 mg, 0.34mmol). The resulting mixture was heated to 100° C. overnight under N₂.After cooling to room temperature, the mixture was diluted with waterand extracted with EtOAc, the organic phase was dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to give a residue, whichwas purified by column chromatography on silica gel (eluted withPE:EA=5:1) to provide the title compound (1.7 g, yield 54%) as a whitesolid.

Example 35 N-(2-chloropyrimidin-4-yl)-6-fluoro-1H-indazol-5-amine

To a mixture of 2,4-dichloropyrimidine (730 mg, 4.89 mmol) and6-fluoro-5-aminoindazole (740 mg, 4.89 mmol) in anhydrous ethanol (15mL) was added Na₂CO₃ (1.56 g, 14.7 mmol). The resulting mixture washeated at 80° C. overnight under N₂. After cooling to room temperature,the mixture was diluted with water and extracted with EtOAc, the organicphase was dried over anhydrous Na₂SO₄ and concentrated under reducedpressure to give a residue, which was purified by column chromatographyon silica gel (eluted with PE:EA=3:1) to provide the title compound (750mg, yield 58%) as a brown solid.

Example 36 tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-6-fluoro-1H-indazole-1-carboxylate

To a solution of N-(2-chloropyrimidin-4-yl)-6-fluoro-1H-indazol-5-amine(750 mg, 2.85 mmol) in anhydrous DCM (10 mL) was added Boc₂O (1.86 g,8.55 mmol), TEA (864 mg, 8.55 mmol) and DMAP (173 mg, 1.42 mmol). Theresulting mixture was stirred at room temperature for 4 hrs. The mixturewas diluted with water and extracted with DCM. The organic phase wasdried over anhydrous Na₂SO₄, and the solvent was removed in vacuum togive a residue, which was purified by column chromatograph on silica gel(eluted with PE:EA=20:1) to give the title compound (800 g, yield 60%)as a white solid.

Example 37 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-6-fluoro-1H-indazole-1-carboxylate

To a mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-6-fluoro-1H-indazole-1-carboxylate(920 mg, 1.98 mmol) in 1,4-dioxane (16 mL) and water (4 mL) was addedN-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(760 mg, 2.38 mmol), Pd(PPh₃)₄ (115 mg, 0.1 mmol) and CsF (906 mg, 5.96mmol). The resulting mixture was heated at 100° C. overnight under N₂.After cooling to room temperature, the mixture was diluted with waterand extracted with EtOAc, the organic phase was dried over anhydrousNa₂SO₄ and concentrated under reduced pressure to give a residue, whichwas purified by column chromatography on silica gel (eluted withPE:EA=3:1) to provide the title compound (600 mg, yield 48%) as a yellowsolid.

Example 382-(3-(4-((6-fluoro-1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamidehydrochloride

To a mixture of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-6-fluoro-1H-indazole-1-carboxylate(520 mg, 0.84 mmol) in EtOAc (2 mL) was added HCl/EtOAc (10 mL) andstirred at room temperature overnight. The formed precipitate wasfiltered and washed with EtOAc, dried in vacuo to afford the titlecompound (200 mg, yield 56%) as a yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ1.04 (d, J=6.4 Hz, 6H), 3.88-3.93 (m, 1H), 4.51 (s, 2H), 7.00(b, 1H), 7.22 (d, J=7.6 Hz, 1H), 7.49 (t, J=7.6 Hz, 1H), 7.56 (d, J=10.4Hz, 1H), 7.75-7.79 (m, 2H), 7.93 (d, J=7.2 Hz, 1H), 8.09 (d, J=6.4 Hz,1H), 8.16 (s, 1H), 8.36 (d, J=6.4 Hz, 1H), 11.09 (s, 1H). MS (ES+) m/e457 (M+H)⁺.

Example 39 N-(2-chloro-5-methoxypyrimidin-4-yl)-1H-indazol-5-amine

To the mixture of 2,4-dichloro-5-methoxypyrimidine (3.56 g, 0.02 mol) inEtOH (80 mL) was added 1H-indazol-5-amine (2.66 g, 0.02 mol), and thenDIEA (7.8 g, 0.06 mol). The resulting mixture was stirred at 45° C.overnight. The reaction mixture was cooled to room temperature andfiltered. The cake was rinsed by MTBE and collected to give the titlecompound (4.4 g, yield 81%) as brown solid.

Example 40 tert-butyl5-((tert-butoxycarbonyl)(2-chloro-5-methoxypyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To the mixture ofN-(2-chloro-5-methoxypyrimidin-4-yl)-1H-indazol-5-amine (4.2 g, 15.3mmol) in DCM (50 mL) were added TEA (4.6 g, 45.9 mmol), (Boc)₂O (8.32 g,38.2 mmol), and DMAP (0.2 g). The resulting mixture was stirred at roomtemperature for 1 h and concentrated followed by purification by columnchromatograph to provide the title compound (5.5 g, yield 76%) as lightyellow solid.

Example 41 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)-5-methoxypyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To the solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloro-5-methoxypyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(1.5 g, 3.16 mmol) in the solvents (dioxane:water=10:1, 33 ml) wereadded N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide (1.2 g, 3.79 mmol), (Boc)₂O (1.38 g, 6.32 mmol), CsF(1.4 g, 9.48 mmol), and then Pd(PPh₃)₄ (0.11 g, 0.095 mmol) under N₂.The resulting mixture was stirred at 90° C. for 24 hrs. The mixture waspurified by column chromatograph to give the title compound (0.98 g,yield 49%) as white solid.

Example 422-(3-(4-((1H-indazol-5-yl)amino)-5-hydroxypyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The mixture of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)-5-methoxypyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(1.2 g, 2 mmol) and pyridine hydrochloride (7.8 g) was stirred at 140°C. for 1 hr. The reaction cooled to room temperature. Water (20 mL) wasadded followed by NH₃.H₂O to adjust the pH to 6-7. The mixture wasfiltered. The cake was collected and dried to give the title compound(0.46 g, yield 55%) as gray solid.

Example 432-(3-(4-((1H-indazol-5-yl)amino)-5-(2-(dimethylamino)ethoxy)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideTFA salt

To the mixture of2-(3-(4-((1H-indazol-5-yl)amino)-5-hydroxypyrimidin-2-yl)phenoxy)-N-isopropylacetamide(450 mg, 1.07 mmol) in THF (45 mL) were added 2-(dimethylamino) ethanol(115 mg, 1.29 mmol), and then PPh₃ (563 mg, 2.15 mmol) was added. Theresulting mixture was stirred at room temperature for 0.5 hr. Then DEAD(374 mg, 2.15 mmol) was added. The resulting mixture was heated atreflux overnight. The solvent was removed under reduced pressurefollowed by addition of 10 mL EtOAc and 10 mL water. TFA was added toadjust pH=4-5. The aqueous solution was concentrated and purified byPrep HPLC to give the title compound (0.1 g, yield 10%) as yellow solid.¹H NMR (400 MHz, DMSO-d₆) δ1.07 (d, J=6.4 Hz, 6H), 2.94 (d, J=4.0 Hz,6H), 3.64-3.65 (m, 2H), 3.92-3.94 (m, 1H), 4.48 (s, 2H), 4.52-4.53 (m,2H), 7.02 (dd, J=9.2 and 1.6 Hz, 1H), 7.37 (t, J=7.2 Hz, 1H), 7.60-7.89(m, 5H), 8.12-8.19 (m, 3H), 8.90 (s, 1H), 9.60 (b, 1H). MS (ES+) m/e 490(M+H)⁺.

Example 44 N-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine

A solution of 2,4,6-trichloropyrimidine (3.67 g, 20 mmol),1H-indazol-5-amine (2.66 g, 20 mmol) and TEA (3.03 g, 30 mmol) in EtOH(75 mL) was heated at reflux overnight. After removing the solvent, theresidue was re-crystallized in MeOH to provide the title compound (4.2g, yield 50%) as a solid.

Example 45N-(2-chloro-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A solution of N-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine (4.2 g,15 mmol), 1-methylpiperazine (2.0 g, 20 mmol) and TEA (3.03 g, 30 mmol)in MeOH (75 mL) was refluxed overnight. After removing the solvent, theresidue was re-crystallized in DCM to give the title compound (3 g,yield 58%) as a solid.

Example 46 tert-butyl5-((tert-butoxycarbonyl)(2-chloro-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a solution ofN-(2-chloro-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)-1H-indazol-5-amine(1.5 g, 4.4 mmol) in DCM (20 mL) were added TEA (0.93 g, 9.2 mmol),(Boc)₂O (3 g, 13.9 mmol) and DMAP (1.1 g, 9.2 mmol). The resultingmixture was stirred at room temperature for 3 hrs. After removing thesolvent, the residue was purified by column chromatography on silica gel(eluting with petroleum ether:ethyl acetate=50:1-10:1) to give the title(1.2 g, yield 50%) as a solid.

Example 47 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloro-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(0.600 g, 1.1 mmol), Pd(dppf)Cl₂ (50 mg), CsF (0.501 g, 3.3 mmol) andN-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(0.640 g, 2 mmol) in dioxane/water (10:1, 10 mL) was stirred at 100° C.overnight. After removing the solvent, the residue was purified by HPLCto give the title compound (250 mg, yield 32%).

Example 482-(3-(4-((1H-indazol-5-yl)amino)-6-(4-methylpiperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideTFA salt

A solution of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)-6-(4-methylpiperazin-1-yl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(250 mg, 0.36 mmol) in DCM (10 mL) and TFA (3.0 mL) was stirred at roomtemperature for 3 hrs. The mixture was concentrated in vacuo to providethe title compound (150 mg, yield 70%) as a solid. ¹H NMR (400 MHz,DMSO-d₆) δ1.02 (d, J=6.4 Hz, 6H), 2.79 (s, 3H), 3.04 (b, 2H), 3.20-3.27(m, 2H), 3.47-3.50 (m, 2H), 3.85-3.93 (m, 1H), 4.45 (s, 2H), 4.71-4.75(m, 2H), 6.56 (s, 1H), 7.01 (dd, J=8.4 and 2.4 Hz, 1H), 7.36 (t, J=8.0Hz, 1H), 7.41-7.52 (m, 4H), 7.88 (d, J=7.6 Hz, 1H), 7.98 (s, 2H), 9.53(s, 1H), 10.00 (s, 1H). MS (ES+) m/e 501 (M+H)⁺.

Example 49 N-(2-chloropyrimidin-4-yl)-6-methyl-1H-indazol-5-amine

A solution of 2,4-dichloropyrimidine (0.69 g, 4.6 mmol),6-methyl-1H-indazol-5-ylamine hydrochloride (0.85 g, 4.6 mmol) and TEA(1.4 g, 13.8 mmol) in EtOH (16 mL) was heated at reflux overnight. Thevolatiles were removed to give the crude title compound (1.7 g), whichwas used for the next step reaction without purification.

Example 50 tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-6-methyl-1H-indazole-1-carboxylate

To a solution of N-(2-chloropyrimidin-4-yl)-6-methyl-1H-indazol-5-amine(1.7 g, crude) in DCM (20 mL) were added TEA (0.93 g, 9.2 mmol), (Boc)₂O(3 g, 13.9 mmol) and DMAP (1.1 g, 9.2 mmol). The resulting mixture wasstirred at room temperature for 3 hrs. After removal of the solvent, theresidue was purified by column chromatography on silica gel (elutingwith petroleum ether:ethyl acetate=50:1-10:1) to give the title compound(0.45 g, yield 21.1% for 2 steps) as a solid.

Example 51 tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-6-methyl-1H-indazole-1-carboxylate

A solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-6-methyl-1H-indazole-1-carboxylate(0.45 g, 1.67 mmol), Pd(dppf)Cl₂ (50 mg), Na₂CO₃ (0.35 g, 3.34 mmol) andN-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(0.64 g, 2 mmol) in dioxane/water (10:1, 10 mL) was stirred at 100° C.overnight. After removing the solvent, the residue was purified by HPLCto give the title compound (180 mg, yield 17%).

Example 52N-isopropyl-2-(3-(4-((6-methyl-1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetamideTFA salt

To a solution of tert-butyl5-((tert-butoxycarbonyl)(2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-6-methyl-1H-indazole-1-carboxylate(180 mg, 0.29 mmol) in DCM (10 mL) was added TFA (1.5 mL) was added. Themixture was stirred at room temperature overnight and concentrated invacuo to provide the title compound (170 mg, yield 96%). MS (ES+) m/e417 (M+H)⁺.

Example 53 N-(2-chloropyrimidin-4-yl)-N-methyl-1H-indazol-5-amine

To a solution of 2,4-dichloropyrimidine (1.0 g, 6.7 mmol) in EtOH (20mL) was added (1H-Indazol-5-yl)-methyl-amine (1.0 g, 6.8 mmol) and TEA(2.02 g, 20 mmol). The resulting mixture was refluxed overnight. Thesolvent was removed to give the crude title compound (2.5 g), which wasused for the next step reaction without purification.

Example 54 tert-butyl5-((2-chloropyrimidin-4-yl)(methyl)amino)-1H-indazole-1-carboxylate

To a solution of N-(2-chloropyrimidin-4-yl)-N-methyl-1H-indazol-5-amine(2.5 g, crude) in DCM (50 mL) were added TEA (2.0 g, 20 mmol), (Boc)₂O(4.2 g, 19.2 mmol), and DMAP (1.0 g) was added. The resulting mixturewas stirred at room temperature for 1 hr. The mixture was concentratedin vacuo and purified by column chromatograph to give the title compound(0.95 g, yield 38.8% for 2 steps) as a solid.

Example 55 tert-butyl5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)(methyl)amino)-1H-indazole-1-carboxylate

To a solution of tert-butyl5-((2-chloropyrimidin-4-yl)(methyl)amino)-1H-indazole-1-carboxylate (0.6g, 1.67 mmol) in the solvent (dioxane:water=4:1, 20 mL) were addedN-isopropyl-2-[3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)-phenoxy]-acetamide(0.6 g, 1.88 mmol), (Boc)₂O (1.09 g, 5 mmol), K₃PO₄ (1.06 g, 5 mmol),t-Bu₃P (0.4 g, 2 mmol) and Pd₂(dba)₃ (0.1 g) under N₂. The resultingmixture was stirred at 100° C. for 24 hrs and concentrated in vacuo toprovide the crude material which was carried out for the next stepreaction without further purification.

Example 562-(3-(4-((1H-indazol-5-yl)(methyl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideTFA salt

To tert-butyl5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)(methyl)amino)-1H-indazole-1-carboxylateas above was added HCl/MeOH (4M, 5 mL) and the mixture was stirred for2.0 hr. The solvent was removed in vacuo and the residue was purified byHPLC to give the title compound (120 mg) as a TFA salt. ¹H NMR (400 MHz,CD3OD) δ1.20 (d, J=6.8 Hz, 6H), 3.85 (s, 3H), 4.07-4.13 (m, 1H), 4.62(s, 2H), 6.37-6.45 (m, 1H), 7.39 (dd, J=10.4 and 2.0 Hz, 1H), 7.43 (d,J=2.0 Hz, 1H), 7.60 (t, J=7.6 Hz, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.86-7.89(m, 2H), 7.91 (s, 1H), 8.05 (b, 1H), 8.18 (s, 1H). MS (ES+) m/e 417(M+H)⁺.

Example 57 2-chloro-N-(1H-pyrazol-4-yl)pyrimidin-4-amine

The mixture of 2, 4-dichloro-pyrimidine (200 mg, 2.41 mmol),1H-pyrazol-4-ylamine (431 mg, 2.89 mmol), and TEA (730 mg, 7.23 mmol) ini-PrOH (8 mL) was stirred at 50° overnight. After cooling, the reactionmixture was concentrated. The crude product was used directly for thenext step without purification.

Example 58 tert-butyl4-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-pyrazole-1-carboxylate

To a mixture of (2-chloro-pyrimidin-4-yl)-(1H-pyrazol-4-yl)-amine (470mg, 2.41 mmol), TEA (730 mg, 7.23 mmol) and DMAP (607 mg, 4.82 mmol) indry DCM (15 mL), Boc₂O (1040 mg, 4.82 mmol) was added slowly. Thereaction mixture was stirred at room temperature for 3 h, andconcentrated. The residue was purified by column chromatography onsilica gel to give the title compound (200 mg, 0.5 mmol, 21% yield) as awhite solid.

Example 59 tert-butyl4-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)amino)-1H-pyrazole-1-carboxylate

To a mixture of4-[tert-butoxycarbonyl-(2-chloro-pyrimidin-4-yl)-amino]-pyrazole-1-carboxylicacid tert-butyl ester (118.5 mg, 0.3 mmol), N-isopropyl-2-(3-(4, 4, 5,5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) acetamide (134 mg,0.42 mmol), Na₂CO₃ (64 mg, 0.6 mmol), and Boc₂O (130 mg, 0.6 mmol) inEtOH (3 mL) and H₂O (0.3 mL), Pd(dppf)₂Cl₂ (21 mg, 0.03 mmol) was added.The mixture was stirred at 1300 under N₂ protection under microwave for30 minutes. After cooling, the mixture was concentrated. The residue waspurified by flash column chromatograph on silica gel, and then purifiedby P-HPLC to give the title compound (30 mg, 0.066 mmol, 22% yield) as awhite solid.

Example 602-(3-(4-((1H-pyrazol-4-yl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideTFA salt

To a solution of tert-butyl 4-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl) pyrimidin-4-yl) amino)-1H-pyrazole-1-carboxylate (167 mg, 0.369mmol) in DCM (20 mL) TFA (2 mL) was added. The mixture was stirred atroom temperature for 5 hrs, and then concentrated to give the titlecompound (170 mg, 0.364 mmol, 98% yield) as a yellow solid. ¹H NMR (400MHz, CD3OD) δ 8.20-8.18 (d, J=7.2 Hz, 1H), 8.07 (s 2H), 7.80-7.78 (d,2H), 7.64-7.60 (t, J=8.8 Hz, 1H), 7.40-7.40 (dd, J=9.6, 2.4 Hz, 1H),6.91-6.89 (d, J=7.2 Hz, 1H), 4.63 (s, 2H), 4.13-4.09 (m, 1H) 1.18-1.16(d, J=6.4 Hz 6H). MS (ES+) m/e 353 (M+H)⁺.

Example 61 5-((2-chloropyrimidin-4-yl)oxy)-1H-indazole

The mixture of 2, 4-dichloro-pyrimidine (184 mg, 1.232 mmol),1H-indazol-5-ol (150 mg, 1.12 mmol), and TEA (340 mg, 3.36 mmol) in EtOH(5 mL) was stirred at 800 overnight. After cooling, the reaction mixturewas concentrated. The crude product was used directly for the next stepwithout purification.

Example 62 tert-butyl5-((2-chloropyrimidin-4-yl)oxy)-1H-indazole-1-carboxylate

To a stirred mixture of 5-((2-chloropyrimidin-4-yl)oxy)-1H-indazole (275mg, 1.12 mmol), TEA (340 mg, 3.36 mmol) and DMAP (28 mg, 0.224 mmol) indry DCM (5 mL), Boc₂O (484 mg, 2.24 mmol) was added slowly. The reactionmixture was stirred at room temperature for 2 hrs, and thenconcentrated. The residue was purified by column chromatography onsilica gel to give the title compound (200 mg, 0.57 mmol, 50% yield) asa white solid.

Example 63 tert-butyl5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl)pyrimidin-4-yl)oxy)-1H-indazole-1-carboxylate

To a mixture of tetr-butyl-5-((2-chloropyrimidin-4-yl)oxy)-1H-indazole-1-carboxylate (104 mg, 0.3 mmol), N-isopropyl-2-(3-(4,4, 5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) phenoxy) acetamide (134mg, 0.42 mmol), t-Bu₃P (61 mg, 0.3 mmol), K₃PO₄.3H₂O (160 mg, 0.6 mmol),and Boc₂O (130 mg, 0.6 mmol) in dioxane (3 mL) and H₂O (0.4 mL),Pd₂(dba)₃ (27 mg, 0.03 mmol) was added. The mixture was stirred at 800under N₂ protection overnight. After cooling, the mixture wasconcentrated. The residue was purified by reverse-phase HPLC to give thetitle compound (58 mg, 0.115 mmol, 38% yield) as a white solid.

Example 642-(3-(4-((1H-indazol-5-yl)oxy)pyrimidin-2-yl)phenoxy)-N-isopropylacetamideHCl salt

The solution of tert-butyl 5-((2-(3-(2-(isopropylamino)-2-oxoethoxy)phenyl) pyrimidin-4-yl) oxy)-1H-indazole-1-carboxylate (340 mg, 0.675mmol) HCl (g)/EtOAc (40 mL) was stirred at room temperature for 3 hrs,and then concentrated to provide the title compound (272 mg, 0.621 mmol,92% yield) as a white solid. ¹H NMR (400 MHz, CD3OD) δ 8.87-8.85 (d,J=6.4 Hz, 1H), 8.25 (s 1H), 7.78-7.71 (m, 2H), 7.69-7.65 (m, 2H),7.50-7.43 (m, 3H), 7.30-727 (dd, J=8.0, 2.4 Hz, 1H), 4.48 (s, 2H),4.06-4.00 (m, 1H), 1.14-1.12 (d, J=6.4 Hz 6H). MS (ES+) m/e 404 (M+H)⁺.

Example 652-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)-1-(piperazin-1-yl)ethanoneHCl salt

The title compound was prepared using essentially the same proceduredescribed for example 12. ¹H NMR (400 MHz, CD3OD) δ 3.24 (b, 4H), 3.82(b, 4H), 5.00 (s, 2H), 6.93 (b, 1H), 7.33 (dd, J=7.6 and 1.6 Hz, 1H),7.56 (t, J=8.4 Hz, 1H), 7.68 (d, J=8.4 Hz, 2H), 7.79 (b, 2H), 8.15 (s,1H), 8.22 (d, J=7.2 Hz, 2H). (ES+) m/e 430 (M+H)⁺.

Example 662-(3-(4-amino-6-chloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide

Example 672-(3-(6-amino-2-chloropyrimidin-4-yl)phenoxy)-N-isopropylacetamide

To a mixture of 4-amino-2, 6-dichloropyrimidine (1.016 g, 6.72 mmol),N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(2.032 g, 6.37 mmol) and CsF (2.858 g, 18.803 mmol) in 1,4-dioxane (31.2mL) and H₂O (6.3 mL) was added Pd(PPh₃)₄ (0.362 g, 0.313 mmol). Theresulting mixture was stirred at 100° C. overnight under N₂. Aftercooling to room temperature, the mixture was concentrated under reducedpressure to give a residue, which was purified by column chromatographyon silica gel (eluted with PE:EtOAc=2:1) to obtain compound2-(3-(4-amino-6-chloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide (670mg, yield 33%) as a white powder and2-(3-(6-amino-2-chloropyrimidin-4-yl)phenoxy)-N-isopropylacetamide (460mg, yield 22%) as a white powder.

Example 682-(3-(4-amino-6-(4-methylpiperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a solution of compound2-(3-(4-amino-6-chloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide (0.97g, 3.031 mmol) in n-BuOH (15 mL) was added 1-methylpiperazine (1.5 g, 15mmol) and stirred at 120° C. overnight under N₂. After cooling to roomtemperature, the mixture was concentrated under reduced pressure to givea residue, which was purified by column chromatography on silica gel(eluted with EtOAc:MeOH=20:1) to obtain the title compound (460 mg,yield 39%) as a light yellow powder.

Example 692-(3-(6-amino-2-(4-methylpiperazin-1-yl)pyrimidin-4-yl)phenoxy)-N-isopropylacetamide

To a solution of compound2-(3-(6-amino-2-chloropyrimidin-4-yl)phenoxy)-N-isopropylacetamide(0.436 g, 1.363 mmol) in n-BuOH (10 mL) was added 1-methylpiperazine(0.682 g, 6.815 mmol) and stirred at 120° C. overnight under N₂. Aftercooling to room temperature, the mixture was concentrated under reducedpressure to give a residue, which was purified by column chromatographyon silica gel (eluted with EtOAc:MeOH=20:1) to obtain the title compound(0.273 g, yield 52%) as a light yellow powder.

Example 70N-isopropyl-2-(3-(4-(4-methylpiperazin-1-yl)-6-(pyridin-4-ylamino)pyrimidin-2-yl)phenoxy)acetamide

To a mixture of compound2-(3-(4-amino-6-(4-methylpiperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(460 mg, 1.199 mmol), 4-iodopyridine (319.5 mg, 1.559 mmol), Pd₂(dba)₃(109.8 mg, 0.12 mmol) and X-Phos (57 mg, 0.12 mmol) in anhydrous1,4-dioxane (15 mL) was added Cs₂CO₃ (1.17 g, 3.3 mmol). The resultingmixture was heated to 120° C. overnight under N₂. After cooling to roomtemperature, the mixture was diluted with 1,4-dioxane and filteredthrough celite pad. The filtrate was concentrated and the residue waswashed with EtOAc and dried in vacuo to obtain the title compound (173mg, yield 31.3%) as a white solid. ¹H NMR (400 MHz, CD3OD) δ 1.17 (d,J=6.4 Hz, 6H), 2.36 (s, 3H), 2.55 (t, J=5.2 Hz, 4H), 3.72 (t, J=4.8 Hz,4H), 4.06-4.16 (m, 1H), 4.56 (s, 2H), 5.97 (s, 1H), 7.19 (dd, J=8.0 and2.4 Hz, 1H), 7.39 (t, J=8.0 Hz, 1H), 7.78 (d, J=6.4 Hz, 2H), 7.98 (s,1H), 8.02 (d, J=8.0 Hz, 1H), 8.32 (d, J=5.6 Hz, 1H). m/e 462 (M+H)⁺.

Example 71N-isopropyl-2-(3-(2-(4-methylpiperazin-1-yl)-6-(pyridin-4-ylamino)pyrimidin-4-yl)phenoxy)acetamide

To a mixture of compound2-(3-(6-amino-2-(4-methylpiperazin-1-yl)pyrimidin-4-yl)phenoxy)-N-isopropylacetamide(493 mg, 1.286 mmol), 4-iodopyridine (290 mg, 1.415 mmol), Pd₂(dba)₃(117.8 mg, 0.129 mmol) and X-Phos (61.4 mg, 0.129 mmol) in anhydrous1,4-dioxane (15 mL) was added Cs₂CO₃ (1258 mg, 3.858 mmol). Theresulting mixture was heated to 120° C. overnight under N₂. Aftercooling to room temperature, the mixture was diluted with 1,4-dioxaneand filtered through celite pad. The filtrate was concentrated and theresidue was washed with EtOAc and dried in vacuo to obtain the titlecompound (184 mg, yield 31.0%) as a white solid. ¹H NMR (400 MHz, CD3OD)δ 1.17 (d, J=6.4 Hz, 6H), 2.36 (s, 3H), 2.57 (t, J=4.8 Hz, 4H), 3.65 (b,4H), 4.08-4.14 (m, 1H), 4.54 (s, 2H), 6.57 (s, 1H), 7.08 (dd, J=8.0 and2.4 Hz, 1H), 7.39 (t, J=8.4 Hz, 1H), 7.62 (d, J=7.6 Hz, 1H), 7.69-7.73(m, 3H), 8.31 (d, J=6.4 Hz, 1H). m/e 462 (M+H)⁺.

Example 72 tert-Butyl 4-((3-bromophenoxy)methyl)piperidine-1-carboxylate

A solution of tert-butyl 4-(hydroxymethyl)piperidine-1-carboxylate (1.5g, 7.0 mmol), 3-bromophenol (1.5 g, 7.0 mmol) and PPh₃ (2.7 g, 10.5mmol) was stirred in dry THF (30 mL) was stirred at 0° C. under anitrogen atmosphere. To this mixture was added DEAD (1.8 g, 10.5 mmol)dropwise over a period of 5 min, and the reaction was monitored by TLC.After complete disappearance of the starting material, the solvent wasevaporated under reduced pressure and the resulting oil purified bycolumn chromatography (PE/EA, 9/1) to provide the title compound (2.4 gcrude) which was used directly without further purification. m/e 372(M+H)⁺.

Example 73 tert-butyl4-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)piperidine-1-carboxylate

A solution of tert-butyl4-((3-bromophenoxy)methyl)piperidine-1-carboxylate (2.4 g, 6.5 mmol),Pin₂B₂ (2.5 g, 9.7 mmol), Pd(dppf)Cl₂ (250 mg) and potassium acetate(1.9 g, 19.4 mmol) in 50 mL of dioxane was degassed and flushed with N₂,heated at 80° C. for 14 h. The mixture was concentrated to give aresidue, which was diluted with EtOAc (300 μL), filtered, concentratedand was purified by chromatography (EA:PE, 1:10) to give the titlecompound (600 mg, 22%) as a yellow solid. ¹H NMR (300 MHz, CDCl₃) δ 7.38(1H, m), 7.27 (2H, m), 6.96 (1H, m), 3.82 (2H, d, J=6.0 Hz), 2.76 (1H,m), 1.73 (4H, m), 1.84 (4H, m).

Example 74 tert-Butyl5-(tert-butoxycarbonyl(2-(3-((1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-(tert-butoxycarbonyl(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(642 mg, 1.44 mmol), tert-butyl4-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)piperidine-1-carboxylate (600 mg, 1.44 mmol), KOAc (564 mg, 5.76 mmol),Boc₂O (604 mg, 2.88 mmol) and Pd(dppf)Cl₂ (70 mg) in dioxane/water (30mL/3 mL) was degassed and flushed with N₂, heated at 100° C. 16 h. Themixture was concentrated to give a residue, which was diluted with DCM,filtered, concentrated and purified by chromatography (PE/EA, 5/1) togive the title compound (300 mg, crude) as a yellow oil. m/e 701 (M+H)⁺.

Example 75N-(2-(3-(piperidin-4-ylmethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

tert-Butyl-5-(tert-butoxycarbonyl(2-(3-((1-(tert-butoxycarbonyl)piperidin-4-yl)methoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate (250 mg,0.36 mmol) was dissolved in 30 mL HCl/Et₂O (saturated). It was stirredat room temperature over night. The mixture was concentrated to give aresidue, which was diluted with water, and extracted by EA. The waterphase was adjusted to 11 using saturated NaHCO₃ solution. It wasconcentrated and the residue was further purified by preparative TLC toprovide the title compound as a yellow solid (50 mg, 35%). ¹H NMR (300MHz, CD30OD) δ 8.24 (2H, m), 8.04 (1H, s), 7.90 (2H, m), 7.56 (2H, m),7.38 (1H, t, J=6.0 Hz), 7.04 (1H, dd, J=9.0 Hz, J=3.0 Hz), 6.65 (1H, d,J=6.0 Hz), 3.95 (2H, d, J=6.0 Hz), 3.42 (2H, d, J=3.0 Hz), 2.15 (2H, t,J=3.0 Hz), 2.12 (1H, m), 2.07 (2H, q, J=3.0 Hz), 1.69 (2H, q, J=3.0 Hz);m/e 401 (M+H)⁺.

Example 76 tert-Butyl 4-(3-bromophenoxy) piperidine-1-carboxylate

A solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (1.4 g, 7.0mmol), 3-bromophenol (1.2 g, 7.0 mmol) and PPh₃ (2.7 g, 10.4 mmol) wasstirred in dry THF (35 mL) at 0° C. under a nitrogen atmosphere. To thismixture was added DEAD (1.8 g, 10.4 mmol) dropwise over a period of 5min, and the reaction was monitored by TLC. After complete disappearanceof starting material, the solvent was evaporated under reduced pressureand the resulting oil purified by column chromatography (PE/EA, 9/1) toprovide the title compound (1.3 g, 52%). m/e 357 (M+H)⁺.

Example 77tert-Butyl-4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)piperidine-1-carboxylate

A solution of tert-butyl 4-(3-bromophenoxy) piperidine-1-carboxylate(1.3 g, 3.5 mmol), Pin₂B₂ (1.4 g, 5.3 mmol), Pd(dppf)Cl₂ (135 mg) andpotassium acetate (1.0 g, 10.6 mmol) in 20 mL of dioxane was degassed,flushed with N₂, and heated at 80° C. for 14 h. The mixture wasconcentrated to give a residue, which was diluted with EtOAc (200 mL),filtered, concentrated and purified by chromotography (EA:PE, 1:10) togive the title compound (500 mg, 36%) as a yellow solid. ¹H NMR (300MHz, CDCl₃) δ 7.38 (1H, m), 7.32 (1H, m), 7.27 (1H, m), 7.25 (1H, m),7.38 (1H, m), 4.51 (1H, m), 3.69 (2H, m), 3.36 (2H, m), 1.88 (2H, m),1.74 (2H, m), 1.34 (2H, m).

Example 78tert-Butyl-5-(tert-butoxycarbonyl(2-(3-(1-(tert-butoxycarbonyl)piperidin-4-yloxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-(tert-butoxycarbonyl(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(221 mg, 0.5 mmol), tert-butyl4-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)piperidine-1-carboxylate(200 mg, 0.5 mmol), KOAc (196 mg, 2.0 mmol), Boc₂O (210 mg, 1.0 mmol)and Pd(dppf)Cl₂ (40 mg) in dioxane/water (30 mL/3 mL) was degassed andflushed with N₂, heated at 100° C. 16 h. The mixture was concentrated togive a residue, which was diluted with DCM, filtered, concentrated andpurified by chromatography (PE/EA, 5/1) to provide the title compound(180 mg, crude) as a yellow oil. m/e 687 (M+H)⁺.

Example 79N-(2-(3-(Piperidin-4-yloxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

tert-Butyl-5-(tert-butoxycarbonyl(2-(3-(1-(tert-butoxycarbonyl)piperidin-4-yloxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(170 mg, 0.25 mmol) was dissolved in 30 mL HCl/Et₂O (saturated). It wasstirred at room temperature overnight. The mixture was concentrated togive a residue, which was diluted with water, and extracted by EA. Thewater phase was adjusted to 11 using saturated NaHCO₃ solution. It wasconcentrated and the residue was further purified by preparative TLC toprovide the title compound (20 mg, 21%) as a yellow solid. ¹H NMR (300MHz, CD₃OD) δ 8.26 (1H, d, J=6.0 Hz), 8.18 (1H, s), 8.04 (1H, s), 7.96(1H, s), 7.93 (1H, t, J=3.0 Hz), 7.56 (2H, m), 7.42 (1H, t, J=9.0 Hz),7.14 (1H, dd, J=9.0 Hz, J=3.0 Hz), 6.87 (1H, d, J=6.0 Hz), 5.80 (1H, m),3.38 (2H, m), 3.30 (2H, m), 2.16 (2H, m), 2.11 (2H, m); m/e 387 (M+H)⁺.

Example 80 2-(3-Bromo-4-fluorophenoxy)-N-cyclopropylacetamide

A solution of 2-chloro-N-cyclopropylacetamide (1.7 g, 13.1 mmol),3-bromo-4-fluorophenol (2.5 g, 13.1 mmol) and K₂CO₃ (2.7 g, 19.6 mmol)in 20 mL of acetone was heated at 60° C. for 16 h. The mixture wasfiltered and concentrated to give a residue, which was purified bycolumn chromatography (PE/EA, 3/1) to give the title compound (2.5 g,66%) as a yellow solid. m/e 289 (M+H)⁺.

Example 81N-Cyclopropyl-2-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide

A solution of 2-(3-bromo-4-fluorophenoxy)-N-cyclopropylacetamide (2.5 g,8.7 mmol), Pin₂B₂ (3.3 g, 13.0 mmol), Pd(dppf)Cl₂ (330 mg) and potassiumacetate (2.6 g, 26.0 mmol) in 35 mL of dioxane was degassed and flushedwith N₂, heated at 80° C. for 14 h. The mixture was concentrated to givea residue, which was diluted with EtOAc (200 mL), filtered, concentratedand purified by chromatography (EA:PE, 1:5) to give the title compound(600 mg, 21%) as a yellow solid. m/e 336 (M+H)⁺.

Example 822-(3-(4-(1H-indazol-5-ylamino)pyrimidin-2-yl)-4-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (150 mg, 0.61mmol),N-cyclopropyl-2-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(205 mg, 0.61 mmol), KOAc (240 mg, 2.45 mmol) and Pd(dppf)Cl₂ (60 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. 16 h. The mixture was concentrated to give a residue, which wasdiluted with DCM, filtered, concentrated and purified by chromatography(DCM/MeOH, 20/1) followed by further purification by preparative TLC togive the title compound (35 mg, 14%) as a yellow solid. ¹H NMR (300 MHz,CD₃OD) δ 8.26 (1H, d, J=6.0 Hz), 8.17 (1H, s), 8.03 (1H, s), 7.52 (3H,m), 7.13 (2H, m), 6.70 (1H, d, J=6.0 Hz), 4.53 (2H, s), 2.72 (1H, t,J=3.0 Hz), 0.75 (2H, t, J=3.0 Hz), 0.57 (2H, t, J=3.0 Hz); m/e 419(M+H)⁺.

Example 832-(3-(4-(1H-indazol-5-ylamino)-5-methylpyrimidin-2-yl)-4-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine (209mg, 0.80 mmol),N-cyclopropyl-2-(4-fluoro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(270 mg, 0.80 mmol), KOAc (316 mg, 3.22 mmol) and Pd(dppf)Cl₂ (60 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. 16 h. The mixture was concentrated to give a residue, which wasdiluted with DCM, filtered, concentrated and purified by chromatography(DCM/MeOH, 20/1) followed by further purification by preparative TLC togive the title compound (35 mg, 10%) as a yellow solid. ¹H NMR (300 MHz,CD₃OD) δ 8.16 (1H, s), 8.12 (1H, s), 8.02 (1H, s), 7.70 (1H, dd, J=9.0Hz, J=3.0 Hz), 7.54 (1H, d, J=9.0 Hz), 7.44 (1H, m), 7.10 (2H, m), 4.46(2H, s), 2.69 (1H, m), 2.31 (3H, s), 0.73 (2H, t, J=3.0 Hz), 0.55 (2H,t, J=3.0 Hz); m/e 433 (M+H)⁺.

Example 84 2-(3-Bromo-5-fluorophenoxy)-N-cyclopropylacetamide

A solution of 2-chloro-N-cyclopropylacetamide (2.8 g, 20.9 mmol),3-bromo-5-fluorophenol (4.0 g, 20.9 mmol) and K₂CO₃ (4.3 g, 31.4 mmol)in 40 mL of acetone was heated at 60° C. for 16 h. The mixture wasfiltered and concentrated to give a residue, which was purified bycolumn chromatography (PE/EA, 3/1) to give the title compound (4.3 g,71%) as a yellow solid. m/e 288 (M+H)⁺.

Example 85N-cyclopropyl-2-(3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide

A solution of 2-(3-bromo-5-fluorophenoxy)-N-cyclopropylacetamide (4.3 g,14.9 mmol), Pin₂B₂ (5.7 g, 22.4 mmol), Pd(dppf)Cl₂ (600 mg) andpotassium acetate (4.4 g, 44.8 mmol) in 50 mL of dioxane was degassedand flushed with N₂, heated at 80° C. for 14 h. The mixture wasconcentrated to give a residue, which was diluted with EtOAc (200 mL),filtered, concentrated and purified by chromatography (EA:PE, 1:5) togive the title compound (3.2 g, 64%) as a yellow solid. m/e 336 (M+H)⁺.

Example 862-(3-(4-(1H-indazol-5-ylamino)pyrimidin-2-yl)-5-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (340 mg, 1.4mmol),N-cyclopropyl-2-(3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(650 mg, 1.9 mmol), CsF (835 mg, 5.5 mmol) and Pd(dppf)Cl₂ (200 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. 16 h. The mixture was concentrated to give a residue, which wasdiluted with DCM, filtered, concentrated and purified by chromatography(DCM/MeOH, 20/1) to give the title compound (80 mg, 14%) as a yellowsolid. ¹H NMR (300 MHz, CD₃OD) δ 8.26 (1H, d, J=6.0 Hz), 8.08 (1H, s),8.06 (1H, s), 7.78 (1H, s), 7.65 (1H, dd, J=9.0 Hz, J=3.0 Hz), 7.58 (2H,s), 6.89 (1H, dt, J=9.0 Hz, J=3.0 Hz), 6.66 (1H, d, J=6.0 Hz), 4.58 (2H,s), 2.74 (1H, m), 0.76 (2H, t, J=3.0 Hz), 0.58 (2H, t, J=3.0 Hz); m/e419 (M+H)⁺.

Example 872-(3-(4-(1H-indazol-5-ylamino)-5-methylpyrimidin-2-yl)-5-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine (360mg, 1.4 mmol),N-cyclopropyl-2-(3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(650 mg, 1.9 mmol), CsF (835 mg, 5.5 mmol) and Pd(dppf)Cl₂ (200 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. 16 h. The mixture was concentrated to give a residue, which wasdiluted with DCM, filtered, concentrated and purified by chromatography(DCM/MeOH, 20/1) to give the title compound (60 mg, 10%) as a yellowsolid. ¹H NMR (300 MHz, CD₃OD) δ 8.14 (1H, s), 8.04 (2H, m), 7.70 (2H,m), 7.57 (2H, m), 6.81 (1H, dt, J=9.0 Hz, J=3.0 Hz), 4.51 (2H, s), 2.72(1H, m), 2.29 (3H, s), 0.72 (2H, t, J=3.0 Hz), 0.57 (2H, t, J=3.0 Hz);m/e 433 (M+H)⁺.

Example 88 tert-Butyl 3-((3-bromophenoxy)methyl)piperidine-1-carboxylate

A solution of tert-butyl 3-(hydroxymethyl)piperidine-1-carboxylate (935mg, 4.35 mmol), 3-bromophenol (753 mg, 4.35 mmol) and PPh₃ (1.71 mg,6.53 mmol) was stirred in dry THF (30 mL) at 0° C. under a nitrogenatmosphere. To this mixture was added dropwise DEAD (1.14 g, 6.53 mmol)over a period of 5 min, and the reaction was monitored by TLC. Aftercomplete disappearance of starting material, the mixture was poured toEA (50 mL), washed with brine (3×20 mL), dried over Na₂SO₄ and filtered.The filtrate was evaporated under reduced pressure and the resulting oilwas purified by column chromatography (PE/EA, 10/1) to get the titlecompound (0.8 g, crude). m/e 370 (M+H)⁺.

Example 89 tert-Butyl3-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)piperidine-1-carboxylate

A solution of tert-butyl3-((3-bromophenoxy)methyl)piperidine-1-carboxylate (0.7 g, 1.9 mmol),Pin₂B₂ (0.72 g, 2.8 mmol), Pd(dppf)Cl₂ (154 mg) and potassium acetate(556 mg, 5.67 mmol) in dioxane (50 mL) was degassed and flushed with N₂,heated at 80° C. for 14 h. The mixture was concentrated to give aresidue, which was diluted with EtOAc (100 mL), washed with brine (3×30mL), dried over Na₂SO₄ and filtered. The filtrate was evaporated underreduced pressure and the resulting oil was purified by columnchromatography (EA:PE, 1:5) to give the title compound (0.9 g, crude).m/e 418 (M+H)⁺.

Example 90tert-Butyl-5-(tert-butoxycarbonyl(2-(3-((1-(tert-butoxycarbonyl)piperidin-3-yl)methoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-(tert-butoxycarbonyl(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(240 mg, 0.54 mmol), tert-butyl3-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)piperidine-1-carboxylate (270 mg, 0.65 mmol), CsF (788 mg, 5.4 mmol),Boc₂O (353 mg, 1.62 mmol) and Pd(dppf)Cl₂ (88 mg) in dioxane/water (20mL/2 mL) was degassed and flushed with N₂, heated at 100° C. for 16 h.The mixture was concentrated to give a residue, which was diluted withEtOAc (100 mL), washed with brine (30 mL×3), dried over Na₂SO₄ andfiltered. The filtrate was evaporated under reduced pressure and theresulting oil was purified by column chromatography (DCM:MeOH, 50:1) togive the title compound (0.1 g) as a yellow oil. m/e 701 (M+H)⁺.

Example 91N-(2-(3-(piperidin-3-ylmethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

tert-Butyl-5-(tert-butoxycarbonyl(2-(3-((1-(tert-butoxycarbonyl)piperidin-3-yl)methoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate (100 mg,0.14 mmol) was dissolved in HFIP (3 mL), the solution was stirred at150° C. for 1 h with M.W. The mixture was concentrated to give aresidue, which was purified by pre-TLC (DCM:MeOH, 4:1) to give the titlecompound as a yellow solid (40 mg, 70%). ¹H NMR (400 MHz, CD₃OD) δ 8.26(d, J=8.4 Hz, 1H), 8.21 (s, 1H), 8.07 (s, 1H), 7.91-7.88 (m, 2H),7.61-7.52 (m, 2H), 7.44 (t, J=8.0 Hz, 1H), 7.14-7.12 (m, 1H), 6.74 (d,J=6.4 Hz, 1H), 4.11-4.07 (m, 1H), 3.97-3.93 (m, 1H), 3.58-3.54 (m, 1H),3.41-3.31 (m, 1H), 3.00-2.89 (m, 2H), 2.36-2.27 (m, 1H), 2.04-1.97 (m,1H), 1.85-1.78 (m, 1H), 1.56-1.45 (m, 1H); m/e 401 (M+H)⁺.

Example 92 tert-Butyl 3-(3-bromophenoxy)pyrrolidine-1-carboxylate

A solution of tert-butyl 3-hydroxypyrrolidine-1-carboxylate (1.0 g, 5.8mmol), 3-bromophenol (1.08 g, 5.8 mmol) and PPh₃ (2.28 g, 8.7 mmol) indry THF (35 mL) was stirred at 0° C. under a nitrogen atmosphere. Tothis mixture was added DEAD (1.51 g, 8.7 mmol) dropwise over a period of5 min, and the reaction was monitored by TLC. After completedisappearance of starting material, the mixture was poured to EA (50mL), washed with brine (20 mL×3), dried over Na₂SO₄ and filtered. Thefiltrate was evaporated under reduced pressure and the resulting oil waspurified by column chromatography (PE/EA, 10/1) to afford the titlecompound (0.8 g, crude). m/e 342 (M+H)⁺.

Example 93 tert-Butyl3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)pyrrolidine-1-carboxylate

A solution of tert-butyl 3-(3-bromophenoxy)pyrrolidine-1-carboxylate(0.8 g, 2.3 mmol), Pin₂B₂ (1.91 g, 7.5 mmol), Pd(dppf)Cl₂ (408 mg) andpotassium acetate (1.47 g, 15 mmol) in dioxane (50 mL) was degassed andflushed with N₂, heated at 80° C. for 14 h. The mixture was concentratedto give a residue, which was diluted with EtOAc (100 mL), washed withbrine (3×30 mL), dried over Na₂SO₄ and filtered. The filtrate wasevaporated under reduced pressure and the resulting oil was purified bycolumn chromatography (EA:PE, 1:5) to give the title compound as ayellow oil (450 mg, crude). m/e 390 (M+H)⁺.

Example 94tert-Butyl-5-(tert-butoxycarbonyl(2-(3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yloxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-(tert-butoxycarbonyl(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(466 mg, 1.05 mmol), tert-butyl3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)pyrrolidine-1-carboxylate(488 mg, 1.25 mmol), CsF (1.53 g, 10.5 mmol), Boc₂O (687 mg, 3.15 mmol)and Pd(dppf)Cl₂ (172 mg) in dioxane/water (30 mL/3 mL) was degassed andflushed with N₂, heated at 100° C. for 16 h. The mixture wasconcentrated to give a residue, which was diluted with EtOAc (100 mL),washed with brine (3×30 mL), dried over Na₂SO₄ and filtered. Thefiltrate was evaporated under reduced pressure and the resulting oil waspurified by column chromatography (PE/EA, 5/1) to give the titlecompound as a yellow solid (200 mg, 28.5%). m/e 673 (M+H)⁺.

Example 95N-(2-(3-(pyrrolidin-3-yloxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

tert-Butyl-5-(tert-butoxycarbonyl(2-(3-(1-(tert-butoxycarbonyl)pyrrolidin-3-yloxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(135 mg, 0.25 mmol) was dissolved in 2 mL of con. HCl. It was stirredfor 5 minutes at room temperature. 10 mL of water was added and thenadjust pH 9-10 by IN NaOH. Extracted with EA (30 mL×3), dried overNa₂SO₄ and filtered. The filtrate was evaporated under reduced pressureand the resulting oil was purified by pre-TLC (DCM/MeOH, 5/1) to givethe title compound as a yellow solid (45 mg, 62.5%). ¹H NMR (400 MHz,CD₃OD) δ 8.27 (d, J=6.0 Hz, 1H), 8.17 (s, 1H), 8.05 (s, 1H), 7.98 (d,J=7.6 Hz, 1H), 7.90-7.86 (m, 1H), 7.59-7.53 (m, 2H), 7.44 (t, J=8.0 Hz,1H), 7.11 (dd, J=2.0 Hz, J=2.4 Hz, 1H), 6.67 (d, J=6.0 Hz, 1H),5.28-5.26 (m, 1H), 3.62-3.31 (m, 4H), 2.40-2.25 (m, 2H); m/e 373 (M+H)⁺.

Example 96 2-(5-Bromo-2-fluorophenoxy)-N-cyclopropylacetamide

A solution of 2-chloro-N-cyclopropylacetamide (1.0 g, 7.5 mmol),5-bromo-2-fluorophenol (1.44 g, 7.5 mmol) and K₂CO₃ (1.55 g, 11.25 mmol)in 30 mL of acetone was heated at 60° C. for 16 h. The mixture wasfiltered and concentrated to give a residue, which was purified bycolumn chromatography (PE/EA, 2/1) to give the title as a yellow solid(1.2 g, 55.3%). m/e 288 (M+H)⁺.

Example 97N-cyclopropyl-2-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide

A solution of 2-(5-bromo-2-fluorophenoxy)-N-cyclopropylacetamide (0.83g, 2.88 mmol), Pin₂B₂ (1.1 g, 4.33 mmol), Pd(dppf)Cl₂ (120 mg) andpotassium acetate (0.85 g, 8.64 mmol) in 35 mL of dioxane was degassedand flushed with N₂, heated at 80° C. for 14 h. The mixture wasconcentrated to give a residue, which was diluted with EtOAc (100 mL),washed with brine (3×30 mL), dried over Na₂SO₄ and filtered. Thefiltrate was evaporated under reduced pressure and the resulting oil waspurified by column chromatography (PE/EA, 2/1) to give the titlecompound as an oil (600 mg, 21%). m/e 336 (M+H)⁺.

Example 982-(5-(4-(1H-indazol-5-ylamino)pyrimidin-2-yl)-2-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (147 mg, 0.6mmol),N-cyclopropyl-2-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(300 mg, 0.89 mmol), KOAc (235 mg, 2.4 mmol) and Pd(dppf)Cl₂ (70 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. for 16 h. The mixture was concentrated to give a residue, whichwas diluted with DCM (30 mL) and filtered. The filtrate was concentratedand purified by column chromatography (DCM/MeOH, 10/1) to give the titlecompound as a yellow solid (35 mg, 14%). ¹H NMR (400 MHz, DMSO) δ 13.01(s, 1H), 9.65 (s, 1H), 8.33 (d, J=5.6 Hz, 1H), 8.21-8.20 (m, 1H),8.09-7.96 (m, 3H), 7.62-7.55 (m, 2H), 7.39-7.34 (m, 1H), 6.68 (d, J=6.0Hz, 1H), 4.64 (s, 2H), 2.70-2.65 (m, 1H), 0.63-0.59 (m, 2H), 0.48-0.43(m, 2H); m/e 419 (M+H)⁺.

Example 992-(5-(4-(1H-indazol-5-ylamino)-5-methylpyrimidin-2-yl)-2-fluorophenoxy)-N-cyclopropylacetamide

A mixture of N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine (100mg, 0.38 mmol),N-cyclopropyl-2-(2-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(130 mg, 0.38 mmol), KOAc (151 mg, 1.55 mmol) and Pd(dppf)Cl₂ (50 mg) indioxane/water (20 mL/3 mL) was degassed and flushed with N₂, heated at100° C. for 16 h. The mixture was concentrated to give a residue, whichwas diluted with DCM (30 mL) and filtered. The filtrate was concentratedand purified by column chromatography (DCM/MeOH, 10/1) to give the titlecompound as a yellow solid (10 mg, 2.9%). ¹H NMR (400 MHz, DMSO) δ 13.01(s, 1H), 8.62 (s, 1H), 8.21 (s, 1H), 8.17 (d, J=4.0 Hz, 1H), 8.11-8.09(m, 2H), 7.97-7.94 (m, 1H), 7.85-7.82 (m, 1H), 7.71-7.68 (m, 1H),7.59-7.56 (m, 1H), 7.31-7.26 (m, 1H), 4.58 (s, 2H), 2.68-2.63 (m, 1H),2.25 (s, 3H), 0.66-0.58 (m, 2H), 0.47-0.43 (m, 2H); m/e 433 (M+H)⁺.

Example 100N-(6-chloro-2-(pyrrolidin-1-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine (0.88 g,3.17 mol), pyrrolidine (225 mg, 3.17 mmol) and DIPEA (818 mg, 6.34 mmol)in BuOH (30 mL) was stirred at 120° C. for 12 h. The mixture wasconcentrated to give a residue, which was purified by pre-HPLC to givethe title compound as a white solid (0.8 g, 80%). m/e 315 (M+H)⁺.

Example 101tert-butyl-5-(6-(3-(2-(cyclopropylamino)-2-oxoethoxy)phenyl)-2-(pyrrolidin-1-yl)pyrimidin-4-ylamino)-1H-indazole-1-carboxylate

A mixture ofN-(6-chloro-2-(pyrrolidin-1-yl)pyrimidin-4-yl)-1H-indazol-5-amine (300mg, 0.95 mmol),N-cyclopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(452 mg, 1.43 mmol), CsF (1.38 g, 1.55 mmol) and Pd(dppf)Cl₂ (150 mg) indioxane/water (30 mL/3 mL) was degassed and flushed with N₂, heated at100° C. for 16 h. The mixture was concentrated to give a residue, whichwas diluted with DCM (50 mL) and filtered. The filtrate was concentratedand purified by column chromatography (DCM/MeOH, 10/1) to give the titlecompound as a yellow solid (90 mg, 16%). m/e 570 (M+H)⁺.

Example 1022-(3-(6-(1H-indazol-5-ylamino)-2-(pyrrolidin-1-yl)pyrimidin-4-yl)phenoxy)-N-cyclopropylacetamide

tert-Butyl5-(6-(3-(2-(cyclopropylamino)-2-oxoethoxy)phenyl)-2-(pyrrolidin-1-yl)pyrimidin-4-ylamino)-1H-indazole-1-carboxylate (90 mg, 0.16 mmol) wasdissolved in HFIP (2 mL), the solution was stirred at 150° C. for 1 hwith M.W. The mixture was concentrated to give a residue, which waspurified by pre-TLC (DCM:MeOH, 4:1) to give the title compound as ayellow solid (35 mg, 46.7%). ¹H NMR (400 MHz, DMSO) δ 12.92 (s, 1H),9.27 (s, 1H), 8.32 (s, 1H), 8.19 (d, J=4.4 Hz, 1H), 7.99 (s, 1H), 7.60(s, 1H), 7.56-7.47 (m, 3H), 7.39 (t, J=8.0 Hz, 1H), 7.02 (dd, J=2.0 Hz,J=2.0 Hz, 1H), 6.47 (s, 1H), 4.50 (s, 2H), 3.63-3.60 (m, 4H), 2.73-2.66(m, 1H), 1.95-1.98 (m, 4H), 0.66-0.61 (m, 2H), 0.51-0.47 (m, 2H); m/e470 (M+H)⁺.

Example 103 3-(3-bromophenyl)-N-cyclopropylpropanamide

3-(3-Bromophenyl) propanoic acid (3.0 g, 13.1 mmol) was added to asolution of SOCl2 (10 ml) and was stirred for 2 hours at 70° C. Themixture was concentrated under reduced pressure. The residue wasdissolved in CH2Cl2 (20 ml), then was added dropwise into the mixture ofcyclopropanamine (1.17 g, 19.6 mmol) and triethylamine (4.0 g, 39.3mmol) at 0° C., then the reaction mixture was stirred overnight atambient temperature. The reaction mixture was quenched with IN HCl andthe organic layer was washed with brine, dried, concentrated to residue.The residue was purified by chromatography (PE/EA: 1/1 to 1/2) to givethe title compound as white solid (2.8 g, 79%). ¹H NMR (400 MHz, CDCl₃)δ7.36 (m, 2H), 7.15 (m, 2H), 5.53 (s, 1H), 2.94 (t, 2H), 2.68 (s, 1H),2.41 (t, 2H), 0.77 (m, 2H), 0.44 (m, 2H). m/e 268 (M+H)⁺.

Example 104N-Cyclopropyl-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide

PdCl₂(dppf) (420 mg, 0.5 mmol) was added into the mixture of3-(3-bromophenyl)-N-cyclopropylpropanamide (2.8 g, 10.3 mmol),4,4,4′,4′,5,5,5′,5′-octamethyl-2,2′-bi(1,3,2-dioxaborolane) (3.9 g, 15.5mmol) and KOAc (2.5 g, 25.7 mmol) in dioxane (80 ml). The mixture wasstirred overnight at 100° C. under nitrogen. The reaction mixture wasthen concentrated in vacuo, and the residue was purified bychromatography (PE/EA: 5/1 to 1/1) to give the title compound as anoff-white solid (3.0 g, 92%). ¹H NMR (300 MHz, CDCl₃) δ 7.60-7.67 (m,2H), 7.27-7.33 (m, 2H), 5.47 (s, 1H), 2.92-2.97 (m, 2H), 2.41 (t, 2H),2.04 (s, 1H), 1.34 (s, 12H), 0.70-0.72 (m, 2H), 0.38-0.40 (m, 2H); m/e316 (M+H)⁺.

Example 105 tert-Butyl5-(tert-butoxycarbonyl(2-(3-(3-(cyclopropylamino)-3-oxopropyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

PdCl₂(dppf) (165 mg, 0.21 mmol) was added into the mixture ofN-cyclopropyl-3-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanamide(380 mg, 1.2 mmol), Boc2O (650 mg, 3.0 mmol), CsF (600 mg, 4.0 mmol) andtert-butyl5-(tert-butoxycarbonyl(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(446 mg, 1.0 mmol) in dioxane/H₂O (30 ml, 10/1) under N₂ flow. Themixture was stirred for 24 h at 100° C. under nitrogen. The reactionmixture was extracted with EA (60 ml) and washed with brine, dried,concentrated in vacuo, and the residue was purified by chromatography(PE/EA: 5/1 to 1/5) to give the title compound product (240 mg) as ayellow oil. m/e 599 (M+H)⁺.

Example 1063-(3-(4-(1H-indazol-5-ylamino)pyrimidin-2-yl)phenyl)-N-cyclopropylpropanamide

tert-Butyl-5-(tert-butoxycarbonyl(2-(3-(3-(cyclopropylamino)-3-oxopropyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(220 mg, 0.367 mmol) was added to a mixture of saturated HCl in ethylether (30 mL). The mixture was stirred for 3 hours at ambienttemperature. Then the mixture was filtered and the yellow solid wasadded to HCl (5 ml), then was stirred for 10 minutes and diluted withH2O (50 ml), filtered. The obtained off-white crystals as hydrochloridesalt was added to saturated NaHCO₃ (10 ml) and was stirred for 2 h. Themixture was filtered and the solid was washed with H₂O (10 ml), dried togive the title compound (50 mg, 34%) as an off-white solid. ¹H NMR (300MHz, DMSO) δ13.00 (s, 1H), 9.62 (s, 1H), 8.34-8.32 (m, 3H), 8.22 (m,1H), 8.06 (s, 1H), 7.85 (s, 1H), 7.55 (m, 1H), 7.40 (m, 3H), 6.66 (d,1H), 2.90 (m, 2H), 2.60 (m, 1H), 2.40 (m, 2H), 0.57 (m, 2H), 0.33 (m,2H). m/e 399 (M+H)⁺.

Example 107 2-(3-bromophenylthio)-N-cyclopropylacetamide

A solution of 2-chloro-N-cyclopropylacetamide (1.33 g, 10 mmol),3-bromobenzenethiol (1.6 g, 8.5 mmol) and K₂CO₃ (4.8 g, 35 mmol) in 30mL of acetone was heated at 70° C. overnight. The mixture was filteredand concentrated to give a residue, which was purified by columnchromatography (PE/EA, 1/1) to give the title compound (2.4 g, 96%) as awhite solid. ¹H NMR δ (300 MHz, CDCl3) 7.39 (1H, m), 7.31 (1H, m), 7.14(2H, m), 6.71 (1H, s), 3.58 (2H, s), 2.64-2.77 (1H, m), 0.73-0.84 (2H,m), 0.41 (2H, m); m/e 286 (M+H)⁺.

Example 108N-cyclopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylthio)acetamide

A solution of 2-(3-bromophenylthio)-N-cyclopropylacetamide (2.43 g, 9.1mmol), Pin₂B₂ (3.5 g, 13.7 mmol), Pd(dppf)Cl₂ (730 mg) and potassiumacetate (2.67 g, 27.3 mmol) in 30 mL of dioxane was degassed and flushedwith N₂, heated at 95° C. for 12 h. The mixture was concentrated to givea residue, which was diluted with EtOAc (200 mL), filtered, concentratedand purified by chromatography (EA:PE, 1:1) to give the title compound(2.4 g, 82%) as a yellow oil. m/e 286 (M+H)⁺.

Example 109 tert-Butyl5-(tert-butoxycarbonyl(2-(3-(2-(cyclopropylamino)-2-oxoethylthio)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture ofN-cyclopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylthio)acetamide(380 mg, 1.1 mmol), N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (246mg, 1.0 mmol), CsF (730 mg, 5.0 mmol), Boc₂O (650 mg, 3.0 mmol), andPd(dppf)Cl₂ (1700 mg) in dioxane/water (27 mL/3 mL) was degassed andflushed with N₂, heated at 100° C. 24 h. The mixture was concentrated togive a residue, which was diluted with DCM, filtered, concentrated andpurified by chromatography (DCM/MeOH, 20/1) to give the crude titlecompound (200 mg) as a yellow solid. m/e 617 (M+H)⁺.

Example 1102-(3-(4-(1H-indazol-5-ylamino)pyrimidin-2-yl)phenylthio)-N-cyclopropylacetamide

A mixture of tert-butyl5-(tert-butoxycarbonyl(2-(3-(2-(cyclopropylamino)-2-oxoethylthio)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(209 mg, 0.80 mmol) in con. HCl (3 mL) was stirred for 10 minutesfollowed by addition of ice. The reaction mixture was adjusted to pH 10with NaHCO₃ solution and extracted with CH₂Cl₂/MeOH (1/1, 20 ml).Filtered and the filtrate was concentrated to give a residue, which waspurified by chromatography (DCM/MeOH, 20/1) followed by furtherpurification by pre-TLC to give the title compound (25 mg, 18%) as ayellow solid. ¹H NMR (300 MHz, CD₃OD) δ 8.36 (1H, m), 8.24 (1H, d, J=6Hz), 8.14 (2H, m), 8.05 (1H, s), 7.56 (2H, s), 7.49 (1H, m), 7.41 (1H,d, J=6 Hz), 6.63 (1H, d, J=6 Hz), 3.60 (2H, s), 2.58 (1H, m), 0.61 (2H,m), 0.36 (2H, m); m/e 417 (M+H)⁺.

Example 111 2-(3-(4-aminopyrimidin-2-yl)phenoxy)-N-isopropylacetamide

A mixture of 2-chloropyrimidin-4-amine (0.50 g, 3.8 mmol),N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(1.46 g, 4.6 mmol), CsF (1.75 g, 11.4 mmol), and Pd(PPh₃)₄ (0.2 g, 0.2mmol) in a mixture of dioxane (8 mL) and H₂O (2 mL) was stirred at 100°C. overnight under N₂. After cooling to room temperature, the mixturewas concentrated under reduced pressure to give a residue, which waspurified by column chromatography on silica gel (eluted withPE:EtOAc=1:1) to provide the title compound (400 mg, yield 36%) ascolourless oil.

Example 112N-isopropyl-2-(3-(4-(pyridin-4-ylamino)pyrimidin-2-yl)phenoxy)acetamide

A mixture of compound2-(3-(4-aminopyrimidin-2-yl)phenoxy)-N-isopropylacetamide

(300 mg, 1.1 mmol), 4-bromopyridine (258 mg, 1.3 mmol), Cs₂CO₃ (1026 mg,3.3 mmol), Pd₂(dba)₃ (96 mg, 0.1 mmol), and X-Phos (51 mg, 0.1 mmol) inanhydrous dioxane (30 mL) was stirred at 120° C. overnight under N₂.After cooling to room temperature, the mixture was filtered, thefiltrate was concentrated, the residue was washed with EtOAc then wasfiltered to provide the title compound (200 mg, yield 52%) as whitesolid. ¹H NMR (400 MHz, CD₃OD) δ 10.09 (s, 1H), 8.47-7.92 (m, 8H), 7.45(t, J=7.6 Hz, 1H), 7.12-6.82 (m, 2H), 4.52 (s, 2H), 3.99-3.92 (m, 1H),1.06 (d, J=6.8 Hz, 6H); m/e 364 (M+H)⁺.

Example 113 2-chloro-4-(4-(pyridin-4-yl)piperidin-1-yl)pyrimidine

2, 4-Dichloropyrimide (745 mg, 5 mmol), 1,2,3,4,5,6-hexahydro-[4,4′]bipyridinyl (811 mg, 5 mmol), and TEA (758 mg, 7.5 mmol) in EtOH (15mL) was stirred at reflux overnight. After removing the solvent, theresidue was purified by column chromatography on silica gel (elutingwith petroleum ether:ethyl acetate=5:1-1:1) to give the title compound(500 mg, yield 36.4%) as a white solid.

Example 114N-isopropyl-2-(3-(4-(4-(pyridin-4-yl)piperidin-1-yl)pyrimidin-2-yl)phenoxy)acetamide

A mixture of 2-chloro-4-(4-(pyridin-4-yl)piperidin-1-yl)pyrimidine (500mg, 1.82 mmol), Pd(dppf)₂Cl₂ (50 mg), Na₂CO₃ (579 mg, 5.46 mmol) andN-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(871 mg, 2.73 mmol) in dioxane/water (10:1, 10 mL) was stirred at 100°C. overnight. After removing the solvent, the residue was purified byP-HPLC to give the title compound (300 mg, yield 35.2%) as a solid. ¹HNMR (400 MHz, CD30OD) δ 8.80 (d, J=6.8 Hz, 2H), 8.23 (d, J=7.6 Hz, 1H),8.10 (d, J=6.4 Hz, 2H), 7.83-7.81 (m, 2H), 7.59 (t, J=8.4 Hz, 1H),7.37-7.20 (m, 2H), 5.54 (d, J=13.2 Hz, 1H), 4.62 (s, 2H), 4.52 (d, J=14Hz, 1H), 4.12-4.06 (m, 1H), 3.61-3.47 (m, 4H), 2.26-1.89 (m, 4H), 1.17(d, J=7.6 Hz, 6H); m/e 432 (M+H)⁺.

Example 115 4,6-dichloro-2-iodopyrimidine

To a solution of compound 4,6-dichloropyrimidin-2-amine (39 g, 237.82mmol) in CH3CN (300 mL), CH₂12 (1000 mL) was added then t-BuONO (129.3g, 1.25 mol) was added and the mixture was heated to reflux overnight.The mixture was concentrated under reduced pressure and the residue waspurified by column chromatography to give compound the title compound(30 g, yield 46%) as a yellow solid.

Example 1162-(3-(4,6-dichloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a mixture of compound 4,6-dichloro-2-iodopyrimidine (13.92 g, 50.64mmol),N-isopropyl-2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)acetamide(18 g, 56.39 mmol), Na₂CO₃ (13.88 g, 130.96) in DME (150 mL) and water(50 mL), Pd(PPh₃)₄ (5.04 g, 4.36 mmol) was added and the mixture washeated to reflux overnight under N₂. Then the reaction mixture waspoured into water (100 mL), extracted with EtOAc (150 mL×2) and theorganic phase was washed by brine, dried with Na₂SO₄ and concentratedunder reduced pressure and the residue was purified by columnchromatography to give the title compound (9.05 g, yield 52%) as whitesolid.

Example 1172-(3-(4-((1H-indazol-5-yl)amino)-6-chloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a solution of compound2-(3-(4,6-dichloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide (5.7 g,16.75 mmol) in iPrOH (110 mL), DIPEA (6.5 g, 48.82 mmol) and1H-indazol-5-amine (2.23 g, 17.25 mmol) were added and the reactionmixture was heated to reflux overnight. The reaction mixture wasconcentrated under reduced pressure and the residue was purified bycolumn chromatography to give the title compound (3.22 g, yield 44%). ¹HNMR (400 MHz, DMSO-d₆) δ 13.05 (s, 1H), 9.90 (s, H), 8.09 (b, 2H),7.95-7.89 (m, 3H), 7.59-7.41 (m, 3H), 7.12 (d, J=8.0 Hz, 1H), 6.66 (s,1H), 4.51 (s, 2H), 4.00-3.94 (m, 1H), 1.08 (d, J=6.4 Hz, 6H); m/e 437(M+H)⁺.

Example 1182-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a stirred solution of2-(3-(4-((1H-indazol-5-yl)amino)-6-chloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide(300 mg, 0.687 mmol) in iprOH (20 mL), Et₃N (3 mL), and piperazine (592mg, 6.87 mmol) were added at room temperature. The mixture was stirredovernight at 110° C. Then reaction mixture was concentrated underreduced pressure and the residue was purified by pre-HPLC to provide thetitle compound (114 mg, yield 34%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.91(s, 1H), 9.01 (s, 1H), 8.08 (s, 1H), 8.02 (s, 1H), 7.94-7.89 (m, 3H),7.51-7.35 (m, 3H), 7.04 (dd, J=8.0 and 2.0 Hz, 1H), 5.82 (s, 1H), 4.50(s, 2H), 3.98-3.92 (m, 1H), 3.48 (b, 4H), 2.76 (b, 4H), 1.07 (d, J=6.8Hz, 6H); m/e 487 (M+H)⁺.

Example 1192-(3-(4-((1H-indazol-5-yl)amino)-6-morpholinopyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (112 mg, yield 34%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.93 (s,1H), 9.09 (s, 1H), 8.08 (s, 1H), 8.03 (s, 1H), 7.95-7.91 (m, 3H),7.49-7.38 (m, 3H), 7.05 (d, J=8.0 Hz, 1H), 5.84 (s, 1H), 4.50 (s, 2H),3.94 (b, 1H), 3.69 (s, 4H), 3.52 (s, 4H), 1.06 (d, J=6.8 Hz, 6H); m/e488 (M+H)⁺.

Example 1202-(3-(4-((1H-indazol-5-yl)amino)-6-(4-methyl-1,4-diazepan-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (100 mg, yield 65%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.94 (s,1H), 9.00 (s, 1H), 8.13 (s, 1H), 8.05 (s, 1H), 7.97-7.91 (m, 3H),7.53-7.05 (m, 4H), 5.75 (s, 1H), 4.53 (s, 2H), 4.02-3.62 (m, 5H), 2.64(b, 2H), 2.50 (b, 2H), 2.26 (s, 3H), 1.92 (b, 2H), 1.09 (d, J=6.8 Hz,6H); m/e 515 (M+H)⁺.

Example 1212-(3-(4-((1H-indazol-5-yl)amino)-6-(1,4-diazepan-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (110 mg, yield 32%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.90 (s,1H), 8.96 (s, 1H), 8.11 (s, 1H), 8.02 (s, 1H), 7.92-7.03 (m, 5H), 5.74(s, 1H), 4.50 (s, 2H), 3.96-3.68 (m, 3H), 2.85 (b, 2H), 2.66 (b, 2H),2.31 (b, 2H), 1.77 (b, 2H), 1.07 (d, J=6.8 Hz, 6H); m/e 501 (M+H)⁺.

Example 1222-(3-(4-((1H-indazol-5-yl)amino)-6-(dimethylamino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (101 mg, yield 33%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.90 (s,1H), 9.00 (s, 1H), 8.10 (s, 1H), 8.02 (s, 1H), 7.97-7.88 (m, 3H), 7.48(s, 2H), 7.38 (t, J=8.0 Hz, 1H), 7.03 (d, J=7.2 Hz, 1H), 5.73 (s, 1H),4.50 (s, 2H), 3.98-3.93 (m, 1H), 3.07 (s, 6H), 1.08 (d, J=6.8 Hz, 6H);m/e 446 (M+H)⁺.

Example 1232-(3-(4-((1H-indazol-5-yl)amino)-6-(piperidin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (110 mg, yield 33%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.91 (s,1H), 8.98 (s, 1H), 8.09 (s, 1H), 8.02 (s, 1H), 7.94-7.89 (m, 3H),7.49-7.03 (m, 5H), 5.85 (s, 1H), 5.00 (s, 2H), 3.96-3.93 (m, 1H), 3.58(b, 4H), 1.55 (b, 6H), 1.07 (d, J=6.8 Hz, 6H); m/e 486 (M+H)⁺.

Example 1242-(3-(4-((1H-indazol-5-yl)amino)-6-((2-methoxyethyl)(methyl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (110 mg, yield 33%). ¹H NMR (400 MHz, DMSO-d₆) δ 13.00 (s,1H), 9.07 (s, 1H), 8.15 (s, 1H), 8.08 (s, 1H), 8.05-7.92 (m, 3H), 7.53(s, 1H), 7.42 (t, J=8.0 Hz, 1H), 7.07 (d, J=7.6 Hz, 1H), 5.79 (s, 1H),4.53 (s, 2H), 4.03-3.95 (m, 1H), 2.78 (b, 2H), 3.59-3.56 (m, 2H), 3.29(s, 3H), 3.07 (s, 3H), 1.10 (d, J=6.8 Hz, 1H); m/e 490 (M+H)⁺.

Example 1252-(3-(4-((1H-indazol-5-yl)amino)-6-((2-(dimethylamino)ethyl)(methyl)amino)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

The title compound was synthesized using the same procedure as that for2-(3-(4-((1H-indazol-5-yl)amino)-6-(piperazin-1-yl)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(example 118) (100 mg, yield 29%). ¹H NMR (400 MHz, DMSO-d₆) δ 12.91 (s,1H), 8.99 (s, 1H), 8.09 (s, 1H), 8.02 (s, 1H), 7.96-7.86 (m, 3H),7.49-7.03 (m, 4H), 5.71 (s, 1H), 4.46 (s, 2H), 4.00-3.92 (m, 1H), 3.67(b, 2H), 3.30 (b, 2H), 3.01 (s, 3H), 2.15 (s, 6H), 1.07 (d, J=6.8 Hz,6H); m/e 503 (M+H)⁺.

Example 1262-(3-(4-chloro-6-(2-(dimethylamino)ethoxy)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a solution of compound2-(3-(4,6-dichloropyrimidin-2-yl)phenoxy)-N-isopropylacetamide (1 g, 2.9mmol) in toluene (24 mL) were added NaOH (232 mg, 5.8 mmol) and2-(dimethylamino)ethanol (261 mg, 2.9 mmol). The resulting mixture wasstirred for 3 hrs at 110° C. Then the reaction mixture was diluted withwater and extracted with DCM. The organic layer was dried over Na₂SO₄,filtered and concentrated under reduced pressure to give a residue whichwas purified by column chromatograph on silica gel (eluted withDCM:MeOH=100:1) to give compound the title compound (550 mg, yield 48%)as a solid.

Example 1272-(3-(4-((1H-indazol-5-yl)amino)-6-(2-(dimethylamino)ethoxy)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide

To a solution of compound2-(3-(4-chloro-6-(2-(dimethylamino)ethoxy)pyrimidin-2-yl)phenoxy)-N-isopropylacetamide(100 mg, 0.25 mmol) in EtOH (1 mL) were added 1H-indozal-5-amine (101.5mg, 0.76 mmol) and TFA (0.25 mL). The resulting mixture was heated to80° C. overnight. The mixture was concentrated and purified bychromatography on silica gel column and purified by Prep-TLC again togive the title compound (100 mg, yield 16%) as a light yellow solid. ¹HNMR (400 MHz, MeOD) δ 8.05-7.96 (m, 4H), 7.54-7.49 (m, 2H), 7.40-7.08(m, 2H), 5.94 (s, 1H), 4.55 (s, 4H), 4.14-4.06 (m, 1H), 2.79-2.76 (m,2H), 2.33 (s, 6H), 1.16 (d, J=6.8 Hz, 6H); m/e 490 (M+H)⁺.

Example 128 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylmorpholine-4-carboxylate

To a mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol(500 mg, 2.27 mmol), DMAP (277.3 mg) and Et₃N (450.46 mg, 4.54, mmol) inDCM (10 ml) was added dropwise a solution of morpholine-4-carbonylchloride (339.5 mg, 2.27 mmol) in DCM (10 ml) at 0° C. Water was addedto the mixture and extracted with DCM (40 mL×2). The organic phase wasdried with Na₂SO₄ and concentrated under reduced pressure to give thetitle compound (600 mg, yield 79%) which was used to next step directly.

Example 129 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenylmorpholine-4-carboxylate

To a stirred solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.2243 mmol) in EtOH (3 mL) and H₂O (0.3 ml) were added Na₂CO₃(47.54 mg, 0.4485 mmol), (Boc)₂O (93.29 mg, 0.4485 mmol) and3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenylmorpholine-4-carboxylate (149.44 mg, 0.4485 mmol) at room temperature.The mixture was degassed by budding nitrogen through the solutionPd(PPh₃)₂Cl₂ (15.07 mg, 0.02243 mmol) was added and the mixture washeated under microwave irradiation for 20 minutes at 110° C. The mixturewas dried and concentrated under reduced pressure and the residue waspurified by column chromatograph on silica gel (DCM:MeOH=50:1) to givethe title compound (50 mg, yield 53%). ¹H NMR (400 MHz, DMSO-d₆) δ 13.02(s, 1H), 9.66 (s, 1H), 8.33-8.18 (m, 4H), 7.55-7.51 (m, 3H), 7.25 (d,J=8.0 Hz, 1H), 6.67 (d, J=5.6 Hz, 1H), 3.64 (b, 6H), 3.45 (b, 2H); m/e417 (M+H)⁺.

Example 130 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyldimethylcarbamate

To a mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol(500 mg, 2.23 mmol), DMAP (277.4 mg) and Et₃N (450.46 mg, 4.46, mmol) inDCM (15 mL) was added dropwise a solution of dimethylcarbamic chloride(238.6 mg, 2.23 mmol) in DCM (15 ml) at 0° C. and the reaction mixturewas stirred overnight at room temperature. Water was added to themixture and extracted with DCM (40 mL×2). The organic phase was driedwith Na₂SO₄ and concentrated under reduced pressure to give the titlecompound (500 mg, yield 76%) which was used to next step directly.

Example 131 3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenyldimethylcarbamate

To a stirred solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.2243 mmol) in EtOH (3 mL) and H₂O (0.3 ml) was added Na₂CO₃(47.54 mg, 0.4485 mmol), (Boc)₂O (93.29 mg, 0.4485 mmol) and3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl dimethylcarbamate(130.58 mg, 0.4485 mmol) at room temperature. The mixture was degassedby budding nitrogen through the solution, Pd(PPh₃)₂Cl₂ (15.07 mg,0.02243 mmol) was added and the mixture was heated under microwaveirradiation for 20 min at 110° C. The mixture was concentrated underreduced pressure and the residue was purified by column chromatograph onsilica gel (DCM:MeOH=50:1) to give the title compound (30 mg, yield37%). ¹H NMR (400 MHz, DMSO-d₆) δ 13.02 (s, 1H), 9.66 (s, 1H), 8.34-8.04(m, 5H), 7.55-7.47 (m, 3H), 7.21 (d, J=8.0 Hz, 1H), 6.67 (d, J=5.6 Hz,1H), 3.07 (s, 3H), 2.94 (s, 3H); m/e 375 (M+H)⁺.

Example 1323-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)pyridine

To a solution of compound3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.5 g, 6.8 mmol)in DMF (20 mL) was added NaH (0.82 g, 20.4 mmol) potionwise at 0° C.with stirring. After 30 minutes, compound 3-(chloromethyl)pyridinehydrochloride (1.4 g, 8.9 mmol) was added portionwise at 0° C., and theresulting mixture was allowed to warm to 20° C. and stirred for 16 hrs.It was quenched with water, extracted with EtOAc (100 mL×3), and theextracts were washed with brine, dried over Na₂SO₄, concentrated underreduced pressure, and the residue was purified by chromatography onsilica gel column (eluted with PE:EA=10:1 to 2:1) to give the titlecompound (1 g, yield 50%) as a white solid.

Example 133 tert-butyl1H-indazol-5-yl(2-(3-(pyridin-3-ylmethoxy)phenyl)pyrimidin-4-yl)carbamate

A mixture of compound3-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)pyridine(1 g, 3.2 mmol), compound tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(670 mg, 1.5 mmol), K₂CO₃ (414 mg, 3 mmol) and Pd(dppf)₂Cl₂ (109 mg,0.15 mmol) in dioxane (20 mL) and H₂O (5 mL) was heated at 90° C. for 16hrs under N₂ atmosphere. Then it was concentrated and the residue waspurified by chromatography on silica gel column (eluted withDCM:MeOH=100:1 to 20:1) to give the title compound (370 mg, yield 50%)as a light red oil.

Example 134 tert-butyl1H-indazol-5-yl(2-(3-(pyridin-3-ylmethoxy)phenyl)pyrimidin-4-yl)carbamate

(370 mg, 0.75 mmol) in DCM (5 mL) was added TFA (5 mL), and theresulting solution was stirred at 20° C. for 3 hrs. It was concentratedand the residue was dissolved in DCM/MeOH (10:1, 100 mL), washed withaqueous K₂CO₃ and brine, dried over Na₂SO₄, concentrated and the residuewas purified by chromatography on silica gel column (eluted withDCM:MeOH=100:1 to 20:1) and recrystallized from MeOH to afford the titlecompound (110 mg, yield 37%) as a light yellow solid. ¹H NMR (400 MHz,DMSO-d₆) δ 13.04 (s, 1H), 9.67 (s, 1H), 8.71 (s, 1H), 8.57 (s, 1H), 8.35(d, J=5.6 Hz, 1H), 8.05 (s, 1H), 8.00-7.89 (m, 4H), 7.56-6.68 (m, 6H),5.25 (s, 2H).

13.02 (s, 1H), 9.66 (s, 1H), 8.33-8.18 (m, 4H), 7.55-7.51 (m, 3H), 7.25(d, J=8.0 Hz, 1H), 6.67 (d, J=5.6 Hz, 1H), 3.64 (b, 6H), 3.45 (b, 2H);m/e 417 (M+H)⁺.

Example 1354-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)pyridine

To a solution of compound3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (1.5 g, 6.8 mmol)in DMF (20 mL) was added NaH (0.82 g, 20.4 mmol) potionwise at 0° C.with stirring. After 30 minutes, compound 4-(chloromethyl)pyridine (1.4g, 8.9 mmol) was added portionwise at 0° C., and the resulting mixturewas allowed to warm to 20° C. and stirred for 16 hrs. It was quenchedwith water, extracted with EtOAc (100 mL×3), and the extracts werewashed with brine, dried over Na₂SO₄, concentrated, and the residue waspurified by chromatography on silica gel column (eluted with PE:EA=10:1to 2:1) to give the title compound (1 g, yield 50%) as a white solid.

Example 136N-(2-(3-(pyridin-4-ylmethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of compound4-((3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)methyl)pyridine(139 mg, 0.44 mmol) (6 batches), compound tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.22 mmol), Na₂CO₃ (47 mg, 0.44 mmol), Boc₂O (96 mg, 0.44 mmol)and Pd(PPh₃)₂Cl₂ (15.4 mg, 0.022 mmol) in EtOH (2 mL) and H₂O (0.2 mL)was heated in microwave reactor at 110° C. for 20 minutes under N₂atmosphere. Then it was cooled, diluted with water, extracted with DCM,the extracts were concentrated and the residue was purified bychromatography on silica gel column (eluted with DCM:MeOH=100:1 to 20:1)and recrystallized from MeOH to afford the title compound (110 mg, yield23%) as a light yellow solid. ¹H NMR (400 MHz, MeOD) δ 8.51 (d, J=5.6Hz, 2H), 8.25 (d, J=6.0 Hz, 1H), 8.15 (s, 1H), 8.05-7.94 (m, 3H),7.57-7.16 (m, 6H), 6.65 (d, J=6.0 Hz, 1H), 5.26 (s, 2H)); m/e 395(M+H)⁺.

Example 1372-(3-(2-methoxyethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

To a mixture of 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol(800 mg, 3.636 mmol) in DCM (20 ml) were added KI (1.2 g, 7.273 mmol),K₂CO₃ (1.307 g 10.091 mmol) and 1-bromo-2-methoxyethane (1.01 g, 7.273mmol) at room temperature. The mixture was stirred overnight at 80° C.The mixture was extracted with DCM (30 mL×2) and the organic phase wasdried with Na₂SO₄ and concentrated under reduced pressure and theresidue was purified by column chromatography to give the title compound(550 mg, yield 50%).

Example 138N-(2-(3-(2-methoxyethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

To a stirred solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.2243 mmol) in EtOH (3 mL) and H₂O (0.3 ml) was added Na₂CO₃(47.54 mg, 0.4485 mmol), (Boc)₂O (93.29 mg, 0.4485 mmol) and2-(3-(2-methoxyethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(124.76 mg, 0.4485 mmol) at room temperature. The mixture was degassedby budding nitrogen through the solution, then Pd(PPh₃)₂Cl₂ (15.07 mg,0.02243 mmol) was added and the mixture was heated under microwaveirradiation for 20 minutes at 110° C. The mixture was concentrated underreduced pressure and the residue was purified by column chromatograph onsilica gel (DCM: MeOH=50:1) to give the title compound (40 mg, yield49%). ¹H NMR (400 MHz, DMSO-d₆) δ 13.01 (s, 1H), 9.61 (s, 1H), 8.32 (d,J=5.6 Hz, 1H), 8.21 (s, 1H), 8.01-7.91 (m, 3H), 7.55-7.36 (m, 3H), 7.04(d, J=7.6 Hz, 1H), 6.65 (d, J=5.6 Hz, 1H), 4.16 (b, 2H), 3.68 (b, 2H);m/e 362 (M+H)⁺.

Example 139 tert-butyl1H-indazol-5-yl(2-(3-methoxyphenyl)pyrimidin-4-yl)carbamate

To a stirred solution of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.2243 mmol) in EtOH (3 mL) and H₂O (0.3 ml) were added Na₂CO₃(47.54 mg, 0.4485 mmol), (Boc)₂O (93.29 mg, 0.4485 mmol) and(3-methoxyphenyl)boronic acid (68.17 mg, 0.4485 mmol) at roomtemperature. The mixture was degassed by budding nitrogen through thesolution, then Pd(PPh₃)₂Cl₂ (15.07 mg, 0.02243 mmol) was added and themixture was heated under microwave irradiation for 20 min at 110° C. Themixture was concentrated under reduced pressure and the residue waspurified by column chromatograph on silica gel to give the titlecompound (80 mg, yield 85%).

Example 140 N-(2-(3-methoxyphenyl)pyrimidin-4-yl)-1H-indazol-5-amine

To a stirred solution of tert-butyl1H-indazol-5-yl(2-(3-methoxyphenyl)pyrimidin-4-yl)carbamate (1 g, 2.39mmol) in DCM (20 mL) was added TFA (10 ml) at room temperature and themixture was stirred overnight at room temperature. Then the mixture wasconcentrated under reduced pressure and the residue was purified bypre_HPLC to give the title compound (200 mg, yield 26%) as white solid.¹H NMR (400 MHz, DMSO-d₆) δ 13.05 (s, 1H), 10.02 (s, 1H), 8.32 (d, J=6.0Hz, 1H), 8.17 (s, 1H), 8.05 (s, 1H), 7.89-7.42 (m, 3H), 7.09 (d, J=7.6Hz, 1H), 6.72 (d, J=6.0 Hz, 1H), 3.83 (s, 3H); m/e 318 (M+H)⁺.

Example 141 4-(2-(3-bromophenoxy)ethyl)morpholine

To a solution of 3-bromophenol (7.8 g, 45.2 mmol) in DMF (120 mL) wasadded 4-(2-chloroethyl)morpholine (8.54 g, 45.2 mmol), K₂CO₃ (414 mg, 3mmol) and KI (7.5 mg, 45.2 mmol), and the resulting mixture was heatedat 20° C. and stirred for 16 hrs. It was quenched with water, extractedwith EtOAc (200 mL×3), and the extracts were washed with brine, driedover Na₂SO₄, concentrated, and the residue was purified bychromatography on silica gel column (eluted with DCM:MeOH=100:1 to 20:1)to give compound the title compound (7 g, yield 54%) as a red liquid.

Example 1424-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)morpholine

A mixture of compound 4-(2-(3-bromophenoxy)ethyl)morpholine (1.1 g, 3.8mmol), BIPN (1.47 mg, 5.8 mmol), KOAc (0.83 mg, 8.5 mmol) andPd(dppf)₂Cl₂ (0.28 mg, 0.38 mmol) in dioxane (15 mL) was heated at 90°C. for 16 hrs under N₂ atmosphere. Then it was concentrated and theresidue was purified by chromatography on silica gel column (eluted withDCM:MeOH=100:1 to 20:1) to give compound the title compound (1 g, yield78%) as a light red oil.

Example 143 tert-butyl1H-indazol-5-yl(2-(3-(2-morpholinoethoxy)phenyl)pyrimidin-4-yl)carbamate

A mixture of compound4-(2-(3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenoxy)ethyl)morpholine(1 g, 3.2 mmol), compound tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(670 mg, 1.5 mmol), K₂CO₃ (414 mg, 3 mmol) and Pd(dppf)₂Cl₂ (109 mg,0.15 mmol) in dioxane (20 mL) and H₂O (5 mL) was heated at 90° C. for 16hrs under N₂ atmosphere. Then it was concentrated and the residue waspurified by chromatography on silica gel column (eluted withDCM:MeOH=100:1 to 20:1) to give the tile compound (380 mg, yield 50%) asa light red oil.

Example 144N-(2-(3-(2-morpholinoethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compound tert-butyl1H-indazol-5-yl(2-(3-(2-morpholinoethoxy)phenyl)pyrimidin-4-yl)carbamate(380 mg, 0.75 mmol) in DCM (5 mL) was added TFA (5 mL), and theresulting solution was stirred at 20° C. for 3 hrs. It was concentratedand the residue was dissolved in DCM/MeOH (10:1, 100 mL), washed withaqueous K₂CO₃ and brine, dried over Na₂SO₄, concentrated and the residuewas purified by chromatography on silica gel column (eluted withDCM:MeOH=80:1 to 15:1) and recrystallized from EtOAc and PE to affordthe title compound (120 mg, yield 39%) as a light yellow solid. ¹H NMR(400 MHz, DMSO-d₆) δ 13.04 (s, 1H), 9.65 (s, 1H), 8.36 (m, 3H), 8.08 (s,1H), 7.94 (s, 1H), 7.57-7.38 (m, 3H), 7.06 (d, J=8.0 Hz, 1H), 6.68 (d,J=5.6 Hz, 1H), 4.17 (t, J=5.6 Hz, 2H), 3.58 (s, 4H), 2.73 (t, J=5.6 Hz,2H), 2.67 (s, 4H); m/e 417 (M+H)⁺.

Example 145 tert-butyl5-((2-(3-acetylphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(100 mg, 0.22 mmol), 3-acetylphenyl)boronic acid (73.7 mg, 0.44 mmol),Na₂CO₃ (47.6 mg, 0.44 mmol), (Boc)₂O (98 mg, 0.44 mmol), Pd(PPh₃)Cl₂ (16mg, 0.022 mmol) in EtOH:H₂O (3.3 mL, 10:1) was heated under microwaveirradiation for 20 min at 110° C. After reaction, it was evaporated, EAand water was added, separated the organic layer, washed with saturatedbrine, dried over Na₂SO₄, concentrated and purified by silica gel togive the title compound (64 mg, yield 67%).

Example 146 tert-butyl5-((2-(3-(1-aminoethyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a stirred solution of tert-butyl5-((2-(3-acetylphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(500 mg, 1.2 mmol), AcONH₄ (924 mg, 12 mmol) in MeOH (10 mL) was addedNaBH₃CN (91 mg, 1.44 mmol), the mixture was stirred 6 hrs at reflux.After reaction, it was evaporated, diluted with water, filtered to givethe title compound as a white solid (300 mg, crude).

Example 147N-(2-(3-(1-aminoethyl)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

A solution of tert-butyl5-((2-(3-(1-aminoethyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(300 mg, crude) in HCl/MeOH (20 mL) was stirred at 40° C. for 6 h. Afterreaction, it was evaporated, then water was added, adjusted the PH to 9with saturated Na₂CO₃, filtered to give the crude product, which waspurified by Pre-HPLC to provide the title compound (100 mg) as a whitesolid. ¹H NMR (400 MHz, DMSO-d₆) δ 13.03 (s, 1H), 9.65 (s, 1H), 8.44 (s,1H), 8.35 (d, J=5.6 Hz, 2H), 8.18 (d, J=7.2 Hz, 1H), 8.05 (s, 1H),7.55-7.40 (m, 4H), 6.67 (d, J=5.6 Hz, 1H), 4.12-4.07 (m, 1H), 2.08 (b,2H), 1.31 (d, J=6.4 Hz, 3H); m/e 331 (M+H)⁺.

Example 148 tert-butyl5-((2-(4-acetylphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(1.5 g, crude), (4-acetylphenyl)boronic acid (1.12 g, 6.8 mmol), Na₂CO₃(721 mg, 6.8 mmol), (Boc)₂O (1.48 g, 6.8 mmol), Pd(PPh₃)Cl₂ (239 mg,0.34 mmol) in EtOH:H₂O (16.5 mL, 10:1) was heated under microwaveirradiation for 20 min at 110° C. After reaction, it was evaporated, EAand water was added, separated the organic layer, washed with saturatedbrine, dried over Na₂SO₄, concentrated to give the title compound (2.6g, crude).

Example 149 tert-butyl5-((2-(4-(1-aminoethyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a stirred solution of tert-butyl5-((2-(4-acetylphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(2.6 g, 6.1 mmol), AcONH₄ (4.7 g, 61 mmol) in MeOH (60 mL) was addedNaBH₃CN (461 mg, 7.32 mmol), the mixture was stirred for 10 h at reflux.After reaction, the solvent was evaporated, then water was added,filtered to give the title compound (2.1 g, crude).

Example 150N-(2-(4-(1-aminoethyl)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

A solution of tert-butyl5-((2-(4-(1-aminoethyl)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(2.1 g, crude) in HCl/MeOH (60 mL) was stirred at 40° C. for 6 h. Afterreaction, the solvent was evaporated, then water was added, adjusted thepH to 9 with saturated Na₂CO₃, filtered to give the crude product, whichwas purified by Pre-HPLC to give the title compound (113.5 mg) as awhite solid. ¹H NMR (400 MHz, DMSO-d₆) δ 13.02 (s, 1H), 9.58 (s, 1H),8.34-8.27 (m, 4H), 8.20 (s, 1H), 8.08 (s, 1H), 7.56 (s, 2H), 7.49 (d,J=6.0 Hz, 1H), 4.05-4.03 (m, 1H), 1.27 (d, J=6.4 Hz, 3H); m/e 331(M+H)⁺.

Example 1512-(3-(2,2-diethoxyethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

To a mixture of compound3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenol (550 mg, 2.5mmol), 2-bromo-1,1-diethoxyethane (985 mg, 5 mmol), Cs₂CO₃ (2.43 g, 7.5mmol) in DMF (25 mL) was added KI (106 mg, 1 mmol), then the mixture wasstirred overnight at 110° C. After reaction, water was added, thenextracted with EA, washed with saturated brine, dried over Na₂SO₄,concentrated to give the title compound (360 mg, crude) as a lightyellow oil.

Example 152 tert-butyl5-((2-(3-(2,2-diethoxyethoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of2-(3-(2,2-diethoxyethoxy)phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane(100 mg, crude), tert-butyl5-((tert-butoxycarbonyl)(2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(70 mg, 0.15 mmol), Na₂CO₃ (65 mg, 0.6 mmol), (Boc)₂O (130 mg, 0.6mmol), Pd(PPh₃)Cl₂ (20 mg, 0.03 mmol) in EtOH:H₂O (4.4 mL, 10:1) washeated under microwave irradiation for 20 min at 110° C. After reaction,it was evaporated, EA and water was added, separated the organic layer,washed with saturated brine, dried over Na₂SO₄, concentrated andpurified by Pre-TLC to give the title compound (30 mg).

Example 1532-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetaldehyde

To a solution of tert-butyl5-((2-(3-(2,2-diethoxyethoxy)phenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(510 mg, 0.98 mmol) in THF (20 mL) was added dropwise 3M HCl (10 mL),then the mixture was stirred at reflux for 7 h. After reaction, it wasevaporated, then water was added, adjusted the pH to 9 with saturatedNa₂CO₃, filtered to give the title compound (420 mg, crude).

Example 154N-(2-(3-(2-(isopropylamino)ethoxy)phenyl)pyrimidin-4-yl)-1H-indazol-5-amine

To a stirred solution of2-(3-(4-((1H-indazol-5-yl)amino)pyrimidin-2-yl)phenoxy)acetaldehyde (420mg, crude), isopropylamine (245 mg, 4.16 mmol) in MeOH (15 mL) was addedNaBH₃CN (131 mg, 2.08 mmol), the mixture was stirred 7 h at reflux.After reaction, it was evaporated and purified by Pre-HPLC to give thetitle compound (90 mg) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ13.02 (s, 1H), 9.65 (s, 1H), 8.34 (d, J=6.0 Hz, 1H), 8.27 (s, 1H), 8.10(s, 1H), 7.94 (d, J=3.2 Hz, 2H), 7.57-7.38 (m, 3H), 7.05 (d, J=8.0 Hz,1H), 6.68 (d, J=6.0 Hz, 1H), 4.106-4.079 (m, 1H), 2.91 (s, 2H),2.80-2.77 (m, 1H), 1.61 (b, 1H), 1.00 (d, J=6.4 Hz, 6H); m/e 389 (M+H)⁺.

Example 155 N-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine

To a stirred solution of 2,4,6-trichloropyrimidine (5.5 g, 30 mmol) inEtOH (100 mL) were added TEA (1.5 g, 45 mmol) and compound1H-indazol-5-amine (3.99 g, 30 mmol) at room temperature. The mixturewas refluxed overnight. After removing the solvent the residue wasre-crystallized in MeOH to give the title compound as a solid (3.4 g,yield 40%).

Example 156 tert-butyl4-(6-((1H-indazol-5-yl)amino)-2-chloropyrimidin-4-yl)piperazine-1-carboxylate

To a stirred solution ofN-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine (1 g, 3.5 mmol) inEtOH (10 mL) was added TEA (1.4 g, 7 mmol), and compound tert-butylpiperazine-1-carboxylate (0.67 g, 3.5 mmol) at room temperature. Themixture was refluxed overnight. After reaction, water was added,separated the organic layer and saturated brine, dried over Na₂SO₄ andconcentrated under reduced pressure to give the title compound (1.2 g)which was used directly for next step reaction without furtherpurification.

Example 157 tert-butyl5-((tert-butoxycarbonyl)(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

To a stirred solution of tert-butyl4-(6-((1H-indazol-5-yl)amino)-2-chloropyrimidin-4-yl)piperazine-1-carboxylate(1.2 g, crude) in DCM (10 mL) was added (Boc)₂O (3 g, 14 mmol), TEA (1.4g, 14 mmol) and DMAP (0.5 g, 3.5 mmol) at room temperature. The mixturewas stirred at room temperature for 30 min. After reaction, water wasadded, separated the organic layer, washed with citric acid monohydrateand saturated brine, dried over Na₂SO₄ and concentrated under reducedpressure. The residue was purified by column chromatograph on silica gelto give the title compound (0.6 g).

Example 158 tert-butyl5-((tert-butoxycarbonyl)(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3-methoxyphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate

A mixture of tert-butyl5-((tert-butoxycarbonyl)(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-chloropyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(600 mg, 0.95 mmol), (3-methoxyphenyl)boronic acid (160 mg, 1.05 mmol),Na₂CO₃ (201 mg, 1.9 mmol), Pd(dppf)Cl₂ (70 mg, 0.095 mmol) indioxane:H₂O (6.6 mL, 10:1) was heated under microwave irradiation for 20min at 140° C. After reaction, it was evaporated, EA and water wasadded, separated the organic layer, washed with saturated brine, driedover Na₂SO₄, concentrated and purified by silica gel to give the titlecompound (250 mg, yield 37.5%).

Example 159N-(2-(3-methoxyphenyl)-6-(piperazin-1-yl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of tert-butyl5-((tert-butoxycarbonyl)(6-(4-(tert-butoxycarbonyl)piperazin-1-yl)-2-(3-methoxyphenyl)pyrimidin-4-yl)amino)-1H-indazole-1-carboxylate(250 mg) in DCM (5 mL) was added TFA (1 mL). The mixture was stirred atroom temperature for 30 min. it was evaporated, then water was added,adjusted the pH to 9 with saturated Na₂CO₃, filtered to give the titlecompound (115 mg, yield 80%) as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ 12.98 (s, 1H), 9.30 (s, 1H), 8.09 (s, 1H), 8.00 (s, 1H), 7.53-7.37 (m,6H), 7.03 (d, J=8.8 Hz, 1H), 6.50 (s, 1H), 3.82 (s, 3H), 3.73 (b, 4H),3.39 (s, 1H), 2.71 (b, 4H); m/e 402 (M+H)⁺.

Example 160N-(2-chloro-6-(2-(dimethylamino)ethoxy)pyrimidin-4-yl)-1H-indazol-5-amine

To a stirred solution ofN-(2,6-dichloropyrimidin-4-yl)-1H-indazol-5-amine (2 g, 7 mmol) in EtOH(20 mL) was added TEA (2.8 g, 7 mmol), and 2-(dimethylamino)ethanol(0.64 g, 7 mmol) at room temperature. The mixture was refluxedovernight. After reaction, water was added, separated the organic layerand saturated brine, dried over Na₂SO₄ and concentrated under reducedpressure to give the title compound (1.5 g). The residue was used intonext step.

Example 161N-(6-(2-(dimethylamino)ethoxy)-2-(3-methoxyphenyl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture ofN-(2-chloro-6-(2-(dimethylamino)ethoxy)pyrimidin-4-yl)-1H-indazol-5-amine(1.5 g, 4.5 mmol), (3-methoxyphenyl)boronic acid (661 mg, 5 mmol),Na₂CO₃ (954 mg, 9 mmol), Pd(dppf)Cl₂ (300 mg, 0.45 mmol) in dioxane:H₂O(22 mL, 10:1) was heated under microwave irradiation for 30 min at 140°C. After reaction, it was evaporated, EA and water was added, separatedthe organic layer, washed with saturated brine, dried over Na₂SO₄ andconcentrated which was purified by Pre-HPLC to give the title compoundas white solid (125 mg). ¹H NMR (400 MHz, DMSO-d₆) δ 13.00 (s, 1H), 9.67(s, 1H), 8.18 (s, 1H), 8.04 (s, 1H), 7.56-7.41 (m, 5H), 7.08-6.86 (m,2H), 4.46-4.43 (m, 1H), 3.83 (s, 3H), 2.66-2.63 (m, 2H), 2.22 (s, 6H);m/e 405 (M+H)⁺.

Example 162

The following compounds were synthesized using the procedures describedabove:

TABLE 1 Example calc Observed No. Structure Formula M + H M + H 163

C₂₂H₂₀N₆O₂ 401 401 164

C₂₃H₂₄N₆O₂ 417 417 165

C₂₃H₂₂N₆O₂ 415 415 166

C₂₄H₂₀N₈O₂ 453 453 167

C₂₅H₂₆N₆O₂ 443 443 168

C₂₃H₂₁F₃N₆O₂ 471 471 169

C₂₃H₂₁F₃N₆O₂ 471 471 170

C₂₃H₂₂N₆O₂ 415 415 171

C₂₄H₂₄N₆O₂ 429 429 172

C₂₄H₂₆N₆O₂ 431 431 173

C₂₅H₂₆N₆O₂ 443 443 174

C₂₃H₂₃N₇O₂ 430 430 175

C₂₃H₂₂N₆O₃ 431 431 176

C₂₃H₁₇N₇O 408 408 177

C₂₂H₂₀N₆O₂ 401 401 178

C₂₂H₂₁N₇O₂ 416 416 179

C₂₃H₂₂N₆O₂ 415 415 180

C₂₄H₂₅N₇O 428 428

Example 181N-(2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine

To a solution of 2,4-dichloropyrimidine (2.0 g, 13.4 mmol) in ethanol(67 mL) was added 5-amino-indazole (1.79 g, 13.4 mmol) and triethylamine(2.81 mL, 20.1 mmol). The mixture was heated at reflux overnight. Thereaction mixture was concentrated and the residue was recrystallizedwith methanol to afford the title compound as a pink solid (3.1 g, 97%).

N-(2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (0.500 g,2.04 mmol), 5-methoxyisoindoline hydrochloride (0.404 g, 2.18 mmol), andK₂CO₃ (0.844 g, 6.11 mmol) in DMF (4.07 mL) was stirred at 115° C.overnight. The mixture was diluted with water and extracted with EtOActwice. The combined organic layers were washed with water and brine,dried over Na₂SO₄, concentrated, and purified by chromatography withMeOH in DCM. The product was further purified by recrystallization inEtOAc to provide the title compound as a light orange solid (215 mg,30%). ¹H NMR (300 MHz, DMSO-d₆) δ 13.15 (s, 1H), 9.45 (s, 1H), 8.52 (s,1H), 8.29 (s, 1H), 8.16 (d, J=5.8 Hz, 1H), 7.81-7.66 (m, 2H), 7.59 (s,1H), 7.25 (d, J=19.2 Hz, 1H), 7.10 (d, J=8.4 Hz, 1H), 6.29 (d, J=5.8 Hz,1H), 5.01 (d, J=16.0 Hz, 4H), 3.99 (s, 3H). MS (ES+) m/e 359 (M+H)⁺.

Example 182 N-(2-(isoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (0.200 g,1.03 mmol), isoindoline (0.140 g, 1.03 mmol), and K₂CO₃ (0.171 g, 1.24mmol) in DMF (6.89 mL) was stirred 80° C. for 4 h followed by at rtovernight. The mixture was diluted with water and extracted with EtOAc.The organic layer was washed with water, brine and dried over MgSO₄ toafford a crude brown oil which was recrystallized from 5% MeOH/DCMfollowed by further recrystallization in EtOH to afford the titlecompound as a grey solid (36.1 mg, 11%). ¹H NMR (300 MHz, DMSO-d₆) δ12.95 (s, 1H), 9.25 (s, 1H), 8.25 (s, 1H), 8.07 (s, 1H), 7.95 (d, J=4.4Hz, 1H), 7.31-7.51 (m, 6H), 6.08 (d, J=4.4 Hz, 1H), 4.82 (d, J=24.7 Hz,4H). MS (ES+) m/e 329 (M+H)⁺.

Example 183N-(5-fluoro-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

N-(2-chloro-5-fluoropyrimidin-4-yl)-1H-indazol-5-amine

To a solution of 2,4-dichloro-5-fluoropyrimidine (0.800 g, 4.55 mmol) inEtOH (40 mL) were added Na₂CO₃ (6.27 g, 45.5 mmol) and compound1H-indazol-5-amine (0.605 g, 4.55 mmol). The resulting mixture wasstirred for 12 h at 100° C. After LCMS showed the reaction wascompleted, the solvent was removed under reduced pressure and theresidue was put into water (50 mL) and extracted with EtOAc (3×100 mL).The organic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure to give a residue which was purified by columnchromatograph on silica gel (eluted with PE:EA=1:1) to give the titlecompound as a solid (120 mg, 9.7%).

N-(5-fluoro-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of N-(2-chloro-5-fluoropyrimidin-4-yl)-1H-indazol-5-amine(300 mg, 1.14 mmol) in acetonitrile (4 mL) were added5-methoxyisoindoline (170 mg, 1.14 mmol) and DIEA (441 mg, 3.42 mmol).The resulting mixture was heated at 90° C. for 12 h. After LCMS showedthe reaction was completed, the mixture was concentrated and dissolvedin methanol and purified by preparative HPLC, and lyophilized to givecompound the title compound as a white solid (52 mg, 12.1%). ¹H NMR (300MHz, DMSO-d₆) δ 13.0 (s, 1H), 9.26 (s, 1H), 8.32 (s, 1H), 8.09 (s, 1H),8.03 (d, J=4.0 Hz, 1H), 7.74 (d, J=8.8 Hz, 1H), 7.50 (d, J=9.2 Hz, 1H),7.30 (b, 1H), 7.00 (b, 1H), 6.86 (dd, J=8.4 and 2.8 Hz, 1H), 4.72 (s,2H), 4.69 (s, 2H), 3.76 (s, 3H). MS (ES+) m/e 377 (M+H)⁺.

Example 184N-(5-chloro-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

N-(2,5-dichloropyrimidin-4-yl)-1H-indazol-5-amine

To a solution of 2,4,5-trichloropyrimidine (1.00 g, 5.49 mmol) in EtOH(20 mL) were added Na₂CO₃ (3.03 g, 28.6 mmol) and compound1H-indazol-5-amine (0.657 g, 4.94 mmol). The resulting mixture wasstirred for 12 h at 15° C. After LCMS showed the reaction was completed,then the solvent was removed under reduced pressure and the residue wasput into water (50 mL) and extracted with EtOAc (3×50 mL). The organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure to give compound the title compound as a solid (800 mg, 52.3%).

N-(5-chloro-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compoundN-(2,5-dichloropyrimidin-4-yl)-1H-indazol-5-amine (281 mg, 1.01 mmol) inacetonitrile (5 mL) were added compound 5-methoxylisoindoline (150 mg,1.01 mmol) and DIEA (391 mg, 3.03 mmol). The resulting mixture washeated at 90° C. for 12 h. After LCMS showed the reaction was completed,the mixture was concentrated and dissolved in methanol and purified bypreparative HPLC, and lyophilized to give compound the title compound asa white solid (115 mg, 29.1%). ¹H NMR (300 MHz, DMSO-d₆) δ 13.00 (s,1H), 8.73 (s, 1H), 8.14 (s, 1H), 8.08 (s, 2H), 7.69 (d, J=8.8 Hz, 1H),7.50 (s, J=8.8 Hz, 1H), 7.27 (b, 1H), 6.97 (b, 1H), 6.84 (d, J=8.4 Hz,1H), 4.67 (t, J=16.4 Hz, 4H), 3.74 (s, 3H). MS (ES+) m/e 393 (M+H)⁺.

Example 185N-(2-(5-methoxyisoindolin-2-yl)-5-methylpyrimidin-4-yl)-1H-indazol-5-amine

N-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compound 2,4-dichloro-5-methylpyrimidine (1.00 g, 6.17mmol) in EtOH (20 mL) were added Na₂CO₃ (3.27 g, 30.9 mmol) and compound1H-indazol-5-amine (0.821 g, 6.17 mmol). The resulting mixture wasstirred for 12 h at 90° C. After LCMS showed the reaction was completed,then the solvent was removed under reduced pressure and the residue wasput into water (50 mL) and extracted with EtOAc (3×50 mL). The organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified by preparative HPLC togive compound the title compound as a solid (400 mg, yield: 25%).

N-(2-(5-methoxyisoindolin-2-yl)-5-methylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compoundN-(2-chloro-5-methylpyrimidin-4-yl)-1H-indazol-5-amine (261 mg, 1.01mmol) in DMF (5 mL) were added compound 5-methoxyisoindoline (150 mg,1.01 mmol) and DIEA (391 mg, 3.03 mmol). The resulting mixture washeated at 110° C. for 12 h. After LCMS showed the reaction wascompleted, the mixture was concentrated and dissolved in methanol andpurified by preparative HPLC, and lyophilized to give compound the titlecompound as a white solid (105 mg, 27.0%). ¹H NMR (300 MHz, DMSO-d₆) δ12.92 (s, 1H), 8.25 (s, 1H), 8.18 (s, 1H), 8.06 (s, 1H), 7.81 (s, 1H),7.72 (d, J=9.2 Hz, 1H), 7.48 (d, J=8.8 Hz, 1H), 7.25 (d, J=8.0 Hz, 1H),6.98 (s, 1H), 6.83 (d, J=8.0 Hz, 1H), 4.70 (s, 2H), 4.65 (s, 2H), 3.75(s, 3H), 2.09 (s, 3H). MS (ES+) m/e 373 (M+H)⁺.

Example 186N-(2-(5-methoxyisoindolin-2-yl)-6-methylpyrimidin-4-yl)-1H-indazol-5-amine

N-(2-chloro-6-methylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compound 2,4-dichloro-6-methylpyrimidine (1.00 g, 6.17mmol) in EtOH (20 mL) were added Na₂CO₃ (3.27 g, 30.9 mmol) and compound1H-indazol-5-amine (0.821 g, 6.17 mmol). The resulting mixture wasstirred for 12 h at 90° C. After LCMS showed the reaction was completed,then the solvent was removed under reduced pressure and the residue wasput into water (50 mL) and extracted with EtOAc (3×50 mL). The organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified by preparative HPLC togive compound the title compound as a solid (400 mg, 25%).

N-(2-(5-methoxyisoindolin-2-yl)-6-methylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution of compoundN-(2-chloro-6-methylpyrimidin-4-yl)-1H-indazol-5-amine (261 mg, 1.01mmol) in DMF (5 mL) were added compound 5-methoxyisoindoline (150 mg,1.01 mmol) and DIEA (391 mg, 3.03 mmol). The resulting mixture washeated at 110° C. for 12 h. After LCMS showed the reaction wascompleted, the mixture was concentrated and dissolved in methanol andpurified by preparative HPLC, and lyophilized to give compound the titlecompound as a white solid (80 mg, 21.4%). ¹H NMR (300 MHz, DMSO-d₆) δ12.91 (s, 1H), 9.08 (s, 1H), 8.25 (s, 1H), 8.06 (s, 1H), 7.47-7.53 (m,2H), 7.29 (b, 1H), 7.00-7.05 (m, 1H), 6.87 (d, J=10.4 Hz, 1H), 5.93 (s,1H), 5.76 (b, 4H), 3.77 (s, 3H), 2.17 (s, 3H). MS (ES+) m/e 373 (M+H)⁺.

Example 187N-(1H-indazol-5-yl)-2-(5-methoxyisoindolin-2-yl)-5,7-dihydrofuro[3,4-d]pyrimidin-4-amine

2-chloro-N-(1H-indazol-5-yl)-5,7-dihydrofuro[3,4-d]pyrimidin-4-amine

To a solution of compound 2,4-dichloro-5,7-dihydrofuro[3,4-d]pyrimidine(1.00 g, 5.30 mmol) in EtOH (30 mL) were added Na₂CO₃ (1.70 g, 15.8mmol) and compound 1H-indazol-5-amine (711 mg, 5.3 mmol). The resultingmixture was stirred for 12 h at 15° C. After LCMS showed the reactionwas completed, the solvent was removed under reduced pressure and theresidue was dissolved in EtOAc (100 mL), washed by water (2×50 mL). Theorganic phase was dried over Na₂SO₄, filtered and concentrated underreduced pressure to give compound the title compound as a solid (800 mg,yield: 38.3%).

N-(1H-indazol-5-yl)-2-(5-methoxyisoindolin-2-yl)-5,7-dihydrofuro[3,4-d]pyrimidin-4-amine

To a solution of compound2-chloro-N-(1H-indazol-5-yl)-5,7-dihydrofuro[3,4-d]pyrimidin-4-amine(346 mg, 1.21 mmol) in acetonitrile (30 mL) were added compound5-methoxyisoindoline (150 mg, 1.00 mmol) and DIEA (273 mg, 2.11 mmol).The resulting mixture was heated at 90° C. for 20 h. After LCMS showedthe reaction was completed, the mixture was concentrated and dissolvedin methanol and purified by preparative HPLC, and lyophilized to givethe title compound (69.0 mg, 17.2%). ¹H NMR (300 MHz, DMSO-d₆) δ 12.90(s, 1H), 8.83 (s, 1H), 8.27 (s, 1H), 8.09 (s, 1H), 7.64 (d, J=8.8 Hz,1H), 7.50 (d, J=8.8 Hz, 1H), 7.00-7.40 (m, 2H), 6.86 (dd, J=8.4 and 2.8Hz, 1H), 4.73-4.89 (m, 8H), 3.77 (s, 3H). MS (ES+) m/e 401 (M+H)⁺.

Example 188N-(4-(5-methoxyisoindolin-2-yl)pyrimidin-2-yl)-1H-indazol-5-amine

2-(2-chloropyrimidin-4-yl)-5-methoxyisoindoline

To a solution of compound 2,4-dichloropyrimidine (298 mg, 2.01 mmol) inEtOH (10 mL) were added Na₂CO₃ (1.11 g, 10.5 mmol) and compound5-methoxyisoindoline (300 mg, 2.01 mmol). The resulting mixture wasstirred for 12 h at 90° C. After LCMS showed the reaction was completed,then the solvent was removed under reduced pressure and the residue wasput into water (50 mL) and extracted with EtOAc (3×50 mL). The organicphase was dried over Na₂SO₄, filtered and concentrated under reducedpressure to give a residue which was purified by preparative TLC to givethe title compound as a solid (250 mg, yield: 47.8%).

N-(4-(5-methoxyisoindolin-2-yl)pyrimidin-2-yl)-1H-indazol-5-amine

To a solution of compound2-(2-chloropyrimidin-4-yl)-5-methoxyisoindoline (250 mg, 0.958 mmol) inDMF (20 mL) were added compound 5-aminoindazole (127 mg, 0.958 mmol) andDIEA (370 mg, 2.87 mmol). The resulting mixture was heated at 110° C.for 20 h. After LCMS showed the reaction was completed, the mixture wasconcentrated and purified by prep-HPLC, and lyophilized to give thetitle compound (51.7 mg, 17%). ¹H NMR (300 MHz, DMSO-d₆) δ 12.83 (s,1H), 9.03 (s, 1H), 8.33 (s, 1H), 8.00 (s, 1H), 7.99 (s, 1H), 7.62 (dd,J=9.2 and 2.0 Hz, 1H), 7.42 (d, J=8.8 Hz, 1H), 7.30 (b, 1H), 7.07 (b,1H), 6.90 (d, J=8.4 Hz, 1H), 6.02 (d, J=6.4 Hz, 1H), 4.83 (d, J=14.4 Hz,2H), 4.65 (d, J=17.6 Hz, 2H). 3.77 (s, 3H). MS (ES+) m/e 359 (M+H)⁺.

Example 189N-(2-(5-fluoroisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (457 mg,1.87 mmol) in DMF (5 mL) were added compound 5-fluoroisoindole (250 mg,1.87 mmol) and DIEA (724 mg, 5.61 mmol). The resulting mixture washeated at 110° C. for 20 h. After LCMS showed the reaction wascompleted, the mixture was concentrated and purified by prep-HPLC, andlyophilized to give the title compound (200 mg, 31.0%). ¹H NMR (300 MHz,DMSO-d₆) δ 12.92 (s, 1H), 9.24 (s, 1H), 8.28 (s, 1H), 8.06 (s, 1H), 7.94(d, J=6.0 Hz, 1H), 7.11-7.53 (m, 5H), 6.08 (d, J=6.4 Hz, 1H), 4.80 (b,4H). MS (ES+) m/e 347 (M+H)⁺.

Example 190N-(2-(5-chloroisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

To a solution of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (384 mg,1.57 mmol) in acetonitrile (5 mL) were added compound5-chloroisoindoline (240 mg, 157 mmol) and DIEA (608 mg, 4.71 mmol). Theresulting mixture was heated at 90° C. for 17 h. After LCMS showed thereaction was completed, the mixture was concentrated and the crude waswashed with MeOH to give the title compound as a brown solid (230 mg,yield: 40.5%). ¹H NMR (300 MHz, DMSO-d₆) δ 13.00 (s, 1H), 9.65 (s, 1H),8.31 (s, 1H), 8.10 (s, 1H), 7.93 (d, J=6.0 Hz, 1H), 7.35-7.57 (m, 5H),6.18 (d, J=6.0 Hz, 1H), 4.82 (b, 4H). MS (ES+) m/e 363 (M+H)⁺.

Example 191N-(2-(6-methoxy-3,4-dihydroisoquinolin-2(1H)-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (100 mg, 0.41mmol), 6-methoxy-1,2,3,4-tetrahydroisoquinoline (81.3 mg, 0.41 mmol),and K₂CO₃ (168 mg, 1.22 mmol) in DMF (1.2 mL) was stirred at 120° C.overnight, cooled to rt, diluted with water, and extracted with EtOAc.The organic layer was concentrated and the residue was purified bychromatography with 1-20% MeOH/DCM to provide the title compound as awhite solid (42 mg, 28%). ¹H NMR (300 MHz, DMSO-d₆) δ 12.96 (s, 1H),9.21 (s, 1H), 8.13 (d, J=1.6 Hz, 1H), 8.04 (t, J=1.1 Hz, 1H), 7.93 (d,J=5.7 Hz, 1H), 7.56-7.41 (m, 2H), 7.15 (d, J=8.2 Hz, 1H), 6.84-6.72 (m,2H), 6.03 (d, J=5.7 Hz, 1H), 4.79 (s, 2H), 3.95 (t, J=5.8 Hz, 2H), 3.73(s, 3H), 2.85 (t, J=6.0 Hz, 2H). MS (ES+) m/e 373 (M+H)⁺.

Example 192N-(2-(6,7-dihydrothieno[3,2-c]pyridin-5(4H)-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (100 mg, 0.41mmol), 4-H thieno[3,2]pyridine (71.5 mg, 0.41 mmol), and K₂CO₃ (168 mg,1.22 mmol) in DMF (1.2 mL) was stirred at 120° C. overnight, cooled tort, diluted with water, and extracted with EtOAc. The organic layer wasconcentrated and the residue was purified by chromatography with 1-20%MeOH/DCM to provide the title compound as a white solid (33 mg, 23%). ¹HNMR (300 MHz, DMSO-d₆) δ 12.97 (s, 1H), 9.23 (s, 1H), 8.12-8.01 (m, 2H),7.93 (d, J=5.7 Hz, 1H), 7.56-7.40 (m, 2H), 7.34 (d, J=5.1 Hz, 1H), 6.97(d, J=5.2 Hz, 1H), 6.04 (d, J=5.7 Hz, 1H), 4.80 (s, 2H), 4.07 (t, J=5.6Hz, 2H), 2.93-2.82 (m, 2H). MS (ES+) m/e 349 (M+H)⁺.

Example 193N-(6-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

N-(6-chloropyrimidin-4-yl)-1H-indazol-5-amine

A mixture of 4,6-dichloropyrimidine (300 mg, 2.01 mmol), tert-butyl5-amino-1H-indazole-1-carboxylate (470 mg, 2.01 mmol),diisopropylethylamine (0.74 mL, 3.03 mmol), and DMF (2.01 mL) wasstirred at 80° C. overnight followed by 120° C. for 4 h. The mixture wascooled to rt, diluted with water, and extracted with EtOAc. The organiclayer was concentrated in vacuo to provide the title compound which wascarried out directly for next step reaction without furtherpurification.

N-(6-(5-methoxyisoindolin-2-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(6-chloropyrimidin-4-yl)-1H-indazol-5-amine (150 mg, 0.61mmol), 5-methoxyisoindoline hydrogen chloride (121 mg, 0.65 mmol), K₂CO₃(253 mg, 1.83 mmol) in DMF (1.22 mL) was stirred at 100° C. for 2 hfollowed by 120° C. for 12 h. The mixture was cooled to rt, diluted withwater, and extracted with EtOAc. The organic layer was concentrated andpurified by chromatography with 0-20% MeOH in DCM to provide the productwhich was further purified by trituration with DCM/Hex to provide puretitle compound (30 mg, 14%). ¹H NMR (300 MHz, DMSO-d₆) δ 12.95 (s, 1H),9.02 (s, 1H), 8.22 (d, J=0.9 Hz, 1H), 8.01 (d, J=2.1 Hz, 2H), 7.55-7.36(m, 2H), 7.31 (d, J=8.4 Hz, 1H), 7.00 (d, J=2.3 Hz, 1H), 6.89 (dd,J=8.4, 2.4 Hz, 1H), 5.74 (d, J=1.1 Hz, 1H), 4.65 (s, 4H), 3.77 (s, 3H).MS (ES+) m/e 359 (M+H)⁺.

Example 194N-(6-(5-methoxyisoindolin-2-yl)-2-methylpyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(6-chloropyrimidin-4-yl)-1H-indazol-5-amine (159 mg, 0.61mmol), 5-methoxyisoindoline hydrogen chloride (121 mg, 0.65 mmol), K₂CO₃(253 mg, 1.83 mmol) in DMF (1.22 mL) was stirred at 100° C. for 2 hfollowed by 120° C. for 12 h. The mixture was cooled to rt, diluted withwater, and extracted with EtOAc. The organic layer was concentrated andpurified by chromatography with 0-20% MeOH in DCM to provide the productwhich was further purified by trituration with DCM/Hex to provide puretitle compound (25 mg, 11%). ¹H NMR (300 MHz, DMSO-d₆) δ 12.95 (s, 1H),8.90 (s, 1H), 7.98 (d, J=15.8 Hz, 2H), 7.54-7.36 (m, 2H), 7.29 (d, J=8.5Hz, 1H), 6.99 (d, J=2.0 Hz, 1H), 6.88 (dd, J=8.3, 2.4 Hz, 1H), 5.60 (s,1H), 4.63 (s, 4H), 3.76 (s, 3H), 2.34 (s, 3H). MS (ES+) m/e 373 (M+H)⁺.

Example 195N-(2-(1H-pyrrolo[3,4-c]pyridin-2(3H)-yl)pyrimidin-4-yl)-1H-indazol-5-amine

A mixture of N-(2-chloropyrimidin-4-yl)-1H-indazol-5-amine (100 mg, 0.41mmol), 2,3-dihydro-1H-pyrrolo[3,4-c]pyridine dihydrogen chloride (102mg, 0.53 mmol), diisopropylethylamine (0.74 mL, 1.22 mmol) in DMF (0.81mL) was stirred at 110° C. overnight. The mixture was cooled to rt,diluted with water, and extracted with EtOAc. The organic layer wasconcentrated and purified by chromatography with 0-20% MeOH in DCM toprovide the title compound (13 mg, 10%) as a slightly yellow solid. ¹HNMR (300 MHz, DMSO-d₆) δ 12.95 (s, 1H), 9.29 (s, 1H), 8.69 (s, 1H), 8.52(d, J=5.0 Hz, 1H), 8.29 (s, 1H), 8.08 (s, 1H), 7.97 (d, J=5.7 Hz, 1H),7.61-7.46 (m, 3H), 6.11 (d, J=5.8 Hz, 1H), 4.88 (s, 4H). MS (ES+) m/e330 (M+H)⁺.

Example 196N-(2-(5-methoxyisoindolin-2-yl)-5,6-dimethylpyrimidin-4-yl)-1H-indazol-5-amine

N-(2-chloro-5,6-dimethylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution of 2,4-dichloro-5,6-dimethylpyrimidine (0.800 g, 4.55mmol) in EtOH (40 mL) were added Na₂CO₃ (2.42 g, 22.8 mmol) and1H-indazol-5-amine (0.605 g, 4.55 mmol). The resulting mixture wasstirred for 12 h at 100° C. The solvent was removed under reducedpressure and the residue was poured into water (50 mL) and extractedwith EtOAc (3×100 mL). The organic phase was dried over Na₂SO₄, filteredand concentrated under reduced pressure to give a residue which waspurified by column chromatograph on silica gel (eluted with PE:EA=1:1)to provide the title compound (120 mg, yield: 9.7%) as a white solid.

N-(2-(5-methoxyisoindolin-2-yl)-5,6-dimethylpyrimidin-4-yl)-1H-indazol-5-amine

To a solution ofN-(2-chloro-5,6-dimethylpyrimidin-4-yl)-1H-indazol-5-amine (1.0 g, 3.66mmol) in CH₃CN (60 mL) were added 5-methoxyisoindoline (545 mg, 3.66mmol) and K₂CO₃ (1.01 g, 7.33 mmol). The resulting mixture was heated at90° C. for 72 h. After LCMS showed the reaction was completed, themixture was concentrated and the crude was washed with MeOH to providethe title compound (423 mg, yield: 30.0%) as a brown solid. ¹H NMR (300MHz, DMSO-d₆) δ 12.95 (s, 1H), 8.08 (s, 1H), 8.01 (d, J=8.8 Hz, 1H),7.30 (d, J=8.0 Hz, 1H), 7.00 (s, 1H), 6.90-6.83 (m, 4H), 4.81 (s, 2H),4.77 (s, 2H), 3.75 (s, 3H), 2.44 (s, 3H), 2.77 (s, 3H). MS (ES+) m/e 387(M+H)⁺.

Example 197N-(1H-indazol-5-yl)-2-(isoindolin-2-yl)-5,7-dihydrofuro[3,4-d]pyrimidin-4-amine

The title compound was synthesized using the same procedure as describedfor the synthesis of Example 7 (KL-00230). ¹H NMR (400 MHz, DMSO-d₆) δ12.94 (s, 1H), 8.82 (s, 1H), 8.25 (s, 1H), 8.06 (s, 1H), 7.64-7.28 (m,6H), 4.87-4.72 (m, 8H). MS (ES+) m/e 371 (M+H)⁺.

Example 198N-(4-(1H-pyrazol-4-yl)phenyl)-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine

N-(4-bromophenyl)-2-chloropyrimidin-4-amine

To a solution of compound N-(4-bromophenyl)-2-chloropyrimidin-4-amine(2.00 g, 13.4 mmol) in EtOH (20 mL) were added Na₂CO₃ (4.26 g, 40.2mmol) and compound 4-bromoaniline (2.3 g, 13.4 mmol). The resultingmixture was stirred for 12 h at 15° C. After LCMS showed the reactionwas completed, then the solvent was removed under reduced pressure andthe residue was put into water (50 mL) and extracted with EtOAc (3×50mL). The organic phase was dried over Na₂SO₄, filtered and concentratedunder reduced pressure to give a residue which was purified by washedwith EA to give compound the title compound (1.20 g, yield: 31.6%) as asolid.

N-(4-bromophenyl)-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine

To a solution of compound N-(4-bromophenyl)-2-chloropyrimidin-4-amine(1.00 g, 3.53 mmol) in acetonitrile (5 mL) were added compound5-methoxyisoindoline (526 mg, 3.53 mmol) and DIEA (1.36 g, 10.6 mmol).The resulting mixture was heated at 90° C. for 17 h. After LCMS showedthe reaction was completed, the mixture was concentrated and the crudewas washed with MeOH to give compound the title compound (600 mg, yield:42.9%) as a brown solid.

2-(5-methoxyisoindolin-2-yl)-N-(4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)phenyl)pyrimidin-4-amine

To the solution of compoundN-(4-bromophenyl)-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine (300 mg,0.758 mol), compound4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole(492 mg, 1.52 mol) and K₂CO₃ (209 mg, 1.52 mol) were dissolved indioxane/H₂O (2 mL/2 mL) and degassed with nitrogen for 10 minutes.Pd(dppf)Cl2 (27.7 mg, 0.0379 mmol) was added and the reaction mixturewas stirred under nitrogen at 85° C. for 17 h. After LCMS showed thereaction was completed, then the mixture was concentrated used to nextstep.

N-(4-(1H-pyrazol-4-yl)phenyl)-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine

To the solution of compound2-(5-methoxyisoindolin-2-yl)-N-(4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazol-4-yl)phenyl)pyrimidin-4-amine(350 mg, 0.0195 mol) was dissolved in TFA (2 mL) and was stirred at 85°C. for 1 h. After LCMS showed the reaction was completed. Then TFA wasremoved and the residue was purified by prep-HPLC to give the titlecompound (53.1 mg, yield: 20.3%) as a brown solid. ¹H NMR (400 MHz,DMSO-d₆) δ 9.55 (s, 1H), 7.99 (s, 1H), 7.94 (d, J=6.0, 1H), 7.76 (d,J=8.4, 1H), 7.57 (d, J=8.4, 1H), 7.31 (s, 1H), 7.00 (s, 1H), 6.89 (d,J=8.4, 1H), 6.13 (d, J=5.6, 1H), 4.77 (m, 4H), 3.76 (s, 3H). MS (ES+)m/e 385 (M+H)⁺.

Example 1992-(5-methoxyisoindolin-2-yl)-N-(pyridin-4-yl)pyrimidin-4-amine

2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine

To a solution compound 2-chloropyrimidin-4-amine (381 mg, 2.95 mmol) andDIEA (692 mg, 5.37 mmol) in CH₃CN (20 mL) was added compound5-methoxyisoindoline (400 mg, 2.68 mmol). Then the reaction was stirredat 90° C. for 16 h. After LCMS showed the reaction was complete. Thecrude product was purified by prep-TLC to give compound the titlecompound (230 mg, yield: 35.4%).

2-(5-methoxyisoindolin-2-yl)-N-(pyridin-4-yl)pyrimidin-4-amine

To a solution of 2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine (270 mg,1.12 mmol) and 4-bromopyridine (193 mg, 1.23 mmol) in dioxane (20 mL)were added Cs₂CO₃ (1.09 g, 3.35 mmol) and stirred under N₂. Then xphos(32.2 mg, 0.0558 mmol) and Pd₂(dba)₃ (51.1 mg, 0.0558 mmol) were addedand the reaction was stirred at 85° C. under N₂ for 16 h. After LCMSshowed the reaction was completed. The crude product was purified byprep-HPLC to give the title compound (96.0 mg, yield: 26.9%). ¹H NMR(400 MHz, DMSO-d₆) δ 9.69 (s, 1H), 8.38 (dd, J=4.8 and 1.6, 2H), 8.07(d, J=5.2, 1H), 7.80 (dd, J=4.8 and 1.2, 2H), 7.33-7.89 (m, 1H),7.04-6.98 (m, 1H), 6.86 (d, J=8.4, 1H), 6.16 (d, J=5.2, 1H), 4.87-4.71(m, 4H), 3.76 (s, 3H). MS (ES+) m/e 320 (M+H)⁺.

Example 2002-(5-methoxyisoindolin-2-yl)-N-(4-(pyridin-4-yl)phenyl)pyrimidin-4-amine

A mixture of compoundN-(4-bromophenyl)-2-(5-methoxyisoindolin-2-yl)pyrimidin-4-amine (297 mg,0.748 mmol), pyridin-4-ylboronic acid (307 mg, 1.496 mmol), K₂CO₃ (206mg, 1.496 mmol), PdCl₂(dppf) (30.5 mg, 0.0374 mmol) and dioxane (20 mL)and H₂O (20 mL) was stirred at 110° C. under N₂ for 12 h. After LCMSshowed the reaction was completed, the mixture was cooled to 28° C.,filtered and the filter cake was washed with MeOH (20 mL), dried to givethe title compound (200 mg, yield: 67.6%) as a brown solid. ¹H NMR (400MHz, DMSO-d₆) δ 9.54 (s, 1H), 8.56 (s, 2H), 8.00-7.69 (m, 7H), 7.29-6.87(m, 3H), 6.14 (s, 1H), 4.78-4.75 (m, 4H), 3.75 (s, 3H). MS (ES+) m/e 396(M+H)⁺.

Example 201 ROCK1 and ROCK2 Compound Selectivity

Dose response curves for Rho-kinase inhibition were derived from aMillipore immuno-based 96 well plate assay (Millipore catalog numberCSA001). Purified active ROCK1 and ROCK2 were obtained from Invitrogen(catalog numbers ROCKI, PV3691 and ROCK2, PV3759). The kit componentsinclude assay plates, which are pre-coated with recombinant MYPT1, whichcontains a specifically phosphorylatable Thr696. The inhibitoryactivities of compounds are measured according to the manufacturesprotocol. Briefly, decreasing concentrations of test compounds or theknown ROCK inhibitor Y-27963, are added, from 50 uM to 0.003 uM toreaction buffer containing 5 mM MgCl2, and 10 mUnits of ROCK1 or ROCK2in assay dilution buffer. This mixture is overlayed into the 96 wellplate and the reaction is initiated with the addition of 2.5 uM ATP. Theassay proceeds at 300 Celsius for 30 minutes with gentle shaking at 120rpm. The assay is terminated by washing of the plate 3 times withTris-buffered saline and tween wash buffer. Anti-phospho-MYPT1 (Thr696)antibody is added to each well to detect the phosphorylated substrateand incubated for 1 hour at room temperature after which HRP conjugatedanti-rabbit IgG secondary is added for 1 hour at room temperature. Afterwashing the assay is developed using a substrate reagent and theabsorbance is read at 450 nm on a Tecan Infinite M1000 reflecting therelative remaining ROCK phosphorylation activity.

Data showing inhibition of ROCK1 and ROCK2, and selectivity of certaincompounds for ROCK2 inhibition, is presented in Table 1.

TABLE 1 IC₅₀ and Ki for ROCK1 and ROCK2 ROCK1 IC₅₀ ROCK2 IC₅₀ ROCK1 KiROCK2 Ki Compound (μM) (μM) (μM) (μM) Ex. 12 1.77 0.54 0.07 0.02 Ex. 2653.45 0.72 2.01 0.03 Ex. 28 6.69 1.96 0.26 0.08 SLx-2119 13.11 1.02 0.500.04 Y-27263 1.13 1.63 0.04 0.06 Ex. 14 4.25 0.17 Ex. 48 0.33 0.47 0.010.02 Ex. 13 22.53 4.64 0.87 0.18Dose response curves for inhibition of ROCK1 vs ROCK2 are shown in FIG.6.

Example 202 ROCK1 and ROCK2 Compound Selectivity

Dose response curves for Rho-kinase inhibition were derived from aInvitrogen Z′-LYTE™ Kinase Assay Kit (Invitrogen catalog number PV3793).Purified active ROCK1 and ROCK2 were obtained from Invitrogen (catalognumbers ROCK1, PV3691 and ROCK2, PV3759). The kit components include acoumarin and fluorescein labeled peptide based on myosin light chain 2(KKRPQRRYSNVF), a proprietary protease containing development reagentand a proprietary Stop buffer used to terminate the developmentreaction. The inhibitory activities of compounds are measured accordingto the manufactures protocol. Briefly, decreasing concentrations of testcompounds or the known ROCK inhibitor Y-27963, are added, from 10 uM to2.56×10⁻⁵ uM to reaction buffer containing 50 mM HEPES pH 7.5, 10 mMMgCl₂, 5 mM EGTA, and 0.05% Brij-35 and of ROCK1 at 0.18 ug/mL or ROCK2at 0.8 ug/mL in assay dilution buffer. This mixture is overlayed into awhite 96-well half area plate and the reaction is initiated with theaddition of 5 uM ATP for ROCK1 or 12 uM ATP for ROCK2. The assayproceeds at room temperature for 1 hour followed by the addition ofdevelopment reagent, and further incubation for 1 hour at roomtemperature. STOP reagent is then added and the reaction and immediatelythe coumarin and fluorescein emission signals are read on a TecanInfinite M1000 fluorescence plate reader (excitation: 400 nm; emission445 and 520 nm, respectively). By comparing the emission ratios of thetest samples against control samples, percent phosphorylation values arecalculated and the concentration of inhibitor that produces ½ inhibitionof kinase activity (IC₅₀) is determined using Prism. Table 2 providesIC₅₀ concentrations for compounds of the above examples. Several of thecompounds also demonstrated activity in a preliminary assay thatmeasured inhibition of myosin light chain phosphorylation (pMLC). Forcompounds marked ND, activity was not determinable under the testconditions employed.

TABLE 2 ROCK Inhibition ROCK2 ROCK1 pMLC ROCK2 ROCK1 pMLC Ex. IC₅₀ IC₅₀inhi- Ex. IC₅₀ IC₅₀ inhi- No. (nM) (nM) bition No. (nM) (nM) bition 14 +91 + 12 ND 75 ND 43 ++ 79 + 48 1000 300 ++ 95 ND 38 + 110 52 82 56 ND 86126 ND 98 112 ND 83 114 ND 87 60 ND 99 74 ND 106 60 ND 102 65 ND 570 >3000 71 + 26 30 3500 117 + 163 80 5900 118 40 6600 + 164 60 5000119 + 165 50 1700 120 166 20 2200 121 + 28 70 2500 122 ND 167 60 3400123 ND 168 30 >10000 124 + 169 >10000 >10000 125 + 170 70 4100 127 ++171 80 7500 129 + 172 120 >10000 131 + 173 30 >10000 134 + 174 191 2800136 + 17 30 1200 138 + 175 500 140 + 176 13 3500 144 + 177 4900 >10000147 >5500 >2000 178 700 4400 150 1000 >3000 179 310 2400 154 40 >1000022 340 10000 159 200 100 ++ 180 380 >10000 161 3200 1000 20 400 >10000

Example 203 ROCK1 and ROCK2 Compound Selectivity

TABLE 3 ROCK Inhibition ROCK2 ROCK1 pMLC ROCK2 ROCK1 pMLC Ex. IC₅₀ IC₅₀inhi- Ex. IC₅₀ IC₅₀ inhi- No. (nM) (nM) bition No. (nM) (nM) bition 18110 893 + 198 2 7066 181 2 434 199 183 30 8340 ++ 200 35 >10000 184 190200 40 >10000 184 140 197 185 50 8750 + 182 60 >10000 + 186 30 2370 +191 700 >10000 + 196 >10 192 1060 >10000 + 187 10 3750 ++ 193130 >10000 + 188 1170 >10000 ND 194 130-180 >10000 + 189 40 2930 ND 195340 >10000 + 190 110 >10000

Example 204 ROCK2 siRNA, but not ROCK1 siRNA Inhibits, IL-17 and IL-21Secretion

To confirm the role of ROCK2 in regulation of IL-17 and IL-21 secretionin human T cells we specifically silenced ROCK1 and ROCK2 expression byRNA interference. Specific ROCK1 and ROCK2 small interfering RNA (siRNA)reduced the protein expression levels by 72% and 84% respectively.Silencing of ROCK2, but not of ROCK1 significantly reduced the IL-17 andIL-21, with minimal effect on IFN-γ secretion in human T cells (FIG. 2).

Example 205 ROCK2 Selective Inhibitor, KD025, Inhibits IL-17/IL-21Secretion and Proliferation in Human CD4⁺ T Cells In Vitro

Activation of resting T cells, resulting in cytokine secretion andproliferation, involves two distinct signals from antigen-presentingcells (APCs), mimicked by co-stimulation of the T cell receptor(TCR)/CD3 complex and the CD28 receptor. Using freshly purified CD4⁺human T cells and stimulatory antibodies against CD3 and CD28 tostimulate IL-17 and IL-21 secretion in response to TCR activation, itwas found that the treatment with ROCK2 selective inhibitor, KD025,significantly inhibited IL-17 and IL-21 secretion in a dose-dependentmanner. Under the same conditions, the inhibition of IFN-γ secretion wasless robust and significant only at high dose (10 μM) of the inhibitor(FIG. 8A). Similarly, any inhibition of IL-2 secretion was observed onlyat the highest concentration (10 μM) of KD025. Consistent with theinhibitory effect on cytokine secretion, the treatment of T cells withKD025 down-regulated their ability to proliferate in response to TCRstimulation in vitro (FIG. 8B).

Example 206 ROCK2 siRNA, but not ROCK1 siRNA Inhibits, IL-17 and IL-21Secretion

To confirm the role of ROCK2 in regulation of IL-17 and IL-21 secretionin human T cells we specifically silenced ROCK1 and ROCK2 expression byRNA interference. Specific ROCK1 and ROCK2 small interfering RNA (siRNA)reduced the protein expression levels by 72% and 84% respectively.Silencing of ROCK2, but not of ROCK1 significantly reduced the IL-17 andIL-21, with minimal effect on IFN-γ secretion in human T cells (FIG. 9).

Example 207 KD025 Inhibits STAT3 Phosphorylation

STAT3 plays a critical role in Th17 differentiation via regulation ofRORγt expression and direct binding to the IL-17 and IL-21 promoters. Inaddition, recent studies have demonstrated that RhoA-dependent STAT3stimulation requires ROCK activity and leads to activation of STAT3phosphorylation on amino acid Y705. Using two different experimentaldesigns, KD025 was demonstrated to significantly down-regulates thephosphorylation of STAT3. In one experiment, T cells were pre-treatedwith KD025 and then stimulation with anti-CD3/CD28 antibodies.Pre-treatment with KD025 resulted in reduced phosphorylation of STAT3(FIG. 11A). In a different experiment, cells were cultured underTh17-skewing conditions for 5 days and then treated with the ROCK2selective inhibitor for 3 hours. STAT3 phosphorylation was reduced bytreatment with the ROCK2 inhibitor (FIG. 11B). In a separate experiment,reduced phosphorylation of STAT3, as well as IFR4 and RORγT, wasconfirmed (FIG. 11C).

Example 208 KD025 Down-Regulates IL-17, IL-21 and IFN-γ Secretion andReduces the Increased Frequency of IFN-γ and IL-17-Expressing Cells inCD4⁺ T Cells from RA Patients

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory diseaseleading to the destruction of joint architecture. The pathogenic eventsthat involved in RA development are not fully understood, although thepivotal role of pro-inflammatory cytokines, such as TNF-α, IL-1β, IFN-β,IL-6 and more recent IL-17 in the induction and maintenance of RApathogenesis is well documented. Moreover, the frequency of Th17 cellsin peripheral blood of RA patients is significantly increased comparedto healthy controls and correlates with disease activity score (DAS).

CD4⁺ T cells were purified from peripheral blood of RA patients atdifferent stages of the disease or from healthy controls and stimulatedusing anti-CD3/CD28 antibodies in presence of KD025 ex vivo. In CD4⁺ Tcells, the ROCK2 selective inhibitor significantly down-regulatedsecretion of IL-17, IL-21 and IFN-γ in response to TCR stimulation in aSTAT3-dependent manner (FIG. 12A). In contrast to healthy controls, thedegree of inhibition of IFN-γ secretion was comparable to inhibitionlevels of IL-17 and IL-21. Among RA patients, inhibition of IFN-γproduction was correlated with disease activity score (DAS). (FIG. 12B).Culture of CD4⁺ T cells from 2 different RA patients in presence ofKD025 significantly reduced the frequencies of both IL-17 andIFN-γ-producing cells as was demonstrated by intracellular staining(FIG. 12C).

Example 209 KD025 Up-Regulates STAT-5 Phosphorylation

The presence of cytokines IL-6, TGF-β, IL-113, IL-23 and antigenstimulation of CD4⁺ T cells leads to induction of IL-17 and IL-21secretion and development of Th17 effector subset. At the same time, thedevelopment and function of Tregs are inhibited. To determine the effectof ROCK2 inhibition on Tregs, freshly purified human CD4⁺ T cells wereactivated using stimulatory antibodies against CD3/CD28 (5 μg/ml), TGF-β(5 ng/ml) and IL-113 (50 ng/ml) for 2 days in presence of 0 μM, 0.5 μM,1 μM, and 2.5 μM concentrations of the selective ROCK2 inhibitor KD025.Western Blot analysis indicates that treatment with KD025 significantlyup-regulates phosphorylation of STAT5 on site Y694 in a dose-dependentmanner. (FIG. 13). Thus, ROCK2 inhibition down-regulates STAT3phosphorylation while STAT5 phosphorylation is increased in human CD4⁺ Tcells that are activated under Th17-skewing conditions.

Example 210 KD025 Increases the Proportion of Foxp3⁺ Cells Among CD4⁺ TCells

Foxp3 is a lineage-specific transcription factor for Tregs and iscrucial to the development and inhibitory function of these cells. STAT5signaling positively regulates the induction and stabilization of Foxp3expression in T cells in vitro and in vivo. Treatment of human CD4⁺ Tcells with selective ROCK2 inhibitor KD025 significantly increased thepercentage of Foxp3⁺ cells after 5 days of Th17 skewing activation invitro. (FIG. 14).

Example 211 KD025 Up-Regulates Treg Inhibition of IL-17 Secretion inCD4⁺CD25⁻ T Cells

Freshly purified CD4⁺CD25⁺ human Tregs were treated with KD025 for 3hours, washed, mixed with CD4⁺CD25⁻ effector T cells at ratio 1:4 and1:9 (Treg:Teff) and activated by using stimulatory antibodies againstCD3 and CD28 for 2 days. KD025-mediated ROCK2 inhibition significantlyincreased the ability of Tregs to suppress IL-17 secretion in CD4⁺CD25⁻effector T cells. (FIG. 15).

Example 212 KD025 Inhibits TGF-β-Induced STAT3 and MLC Phosphorylationin SMAD2/3-Independent Manner

Selective ROCK2 inhibitor KD025 down-regulates TGF-β-induced STAT3 andMLC phosphorylation in a dose-dependent manner, but phosphorylation ofSMAD2/3 remains intact. (FIG. 16). TGF-3 is involved in development andfunction of both Th17 and Treg subsets of T cells. However, when Tregsare regulated in Smad2/3-dependent signaling mechanism, TGF-β regulatesTh17 cells in Smad2/3 independent manner that is considered asnon-canonical TGF-β-induced signaling. Furthermore, it was demonstratedthat ROCK proteins are involved in Smad-independent TGF-β signaling incancer cells.

Example 213 Treatment of Patients with KD025 Inhibits Ex VivoStimulation of IL-17 and IL-21 Production in Isolated PBMCs

Eight patients were administered 120 mg/day KD025 or placebo on days 1and 8-14 of the trial. Peripheral blood mononuclear cells (PBMCs) werecollected at the start and end of the trial and stimulated by plating onimmobilized anti-CD3/CD28 antibodies for 72 hours, and levels of IL-17,IL-21, and IFN-γ were measured. FIG. 17 shows high levels of IL-17 andIL-21 could be induced in PBMCs isolated at day 1 from six of thesubjects. Of those six subjects, five received the ROCK2-selectiveinhibitor KD025, whereas one received placebo. Induction of IL-17secretion was significantly inhibited in PBMCs isolated from four offive patients treated with KD025 (FIG. 17B), and induction of IL-21secretion was significantly inhibited in PBMCs from all five patientstreated with KD025 (FIG. 17A). No effect on IFN-γ was observed.

Inhibition of cytokine induction was then determined in twenty twopatients administered 40 mg/day, 120 mg/day, 240 mg/day, or 320 mg/dayKD025. FIG. 17D-F shows the dose-response relationship for IL-21, IL-17,and IFN-γ secretion, respectively.

Example 214 Reversal of Lung Pathology in Murine Model of Chronic GVHDby Selective ROCK2 Inhibitor

Chronic GVHD induced by allogeneic HCT after a conditioning regimen ofcyclophosphamide and total-body radiation results in pulmonarydysfunction and airway obliteration which leads to bronchiolitisobliterans in patients. KD025 was tested in a previously describedmurine model of chronic GVHD (Srinivasan M et al. Blood 2012) thatinvolves a wide spectrum of target organs, such lung and liver. Briefly,B10.BR recipients were conditioned with cyclophosphamide (Cy) on days −3and −2 (120 mg/kg intra-peritoneally). On day −1, recipients receivedtotal-body irradiation (TBI) by X-ray (8.5 Gy). Donor bone marrow (BM)was T-cell depleted with anti-Thy 1.2 mAb followed by rabbit complement.On day 0, recipients received 10⁷ BM cells with or without allogeneicsplenocytes (10⁶ cells). Weights of individual mice were recorded twiceweekly. Mice (8 per group) were treated intra-peritoneally with KD025(100 mg/kg) on day 28 (manifestation of decreased lung function andpathology) for 1 month on a daily basis. The analysis of pulmonaryfunction was performed on day 28 (before the treatment) and on day 56.Anesthetized mice were weighed, and lung function was assessed bywholebody plethysmography with the Flexivent system (SCIREQ) andanalyzed with Flexivent Version 5.1 software. Survival data wereanalyzed by life-table methods, and actuarial survival rates are shown.Group comparisons were made by log-rank test. The overall experimentaldesign is provided in the table below:

Donor Donor Conditioning Bone Spleno- TBI Treatment Marrow cytes Groupsn Day -1 Days -2 and -3 Recipient Day 0 Day 0 BM 8 850x Cy B10.BR 10⁷none BM + 8 850x Cy B10.BR 10⁷ 10⁶ Vehicle BM + 8 850x Cy B10.BR 10⁷ 10⁶KD025

As shown in FIG. 18, Panel A, in mice transplanted with BM cells andallogeneic splenocytes (“vehicle”), pulmonary function tests (PFTs) onday 56 after transplantation showed an increase in airway resistance,increased lung constriction, increases elastance, and decreasedcompliance. These measures indicate increased stiffness or rigidity ofthe lungs. Treatment with KD025 significantly improved and normalizedpulmonary function, i.e., resulted in decrease in resistance andelastance, and increase in lung compliance compared to vehicle treatedgroup. A decrease in pulmonary function in mice not treated with KD025was noticed as early as 28 days after transplantation. FIG. 19, Panel A,shows that the effect of KD025 on cGVHD is dose dependent. Panels B andC of FIG. 18 and Panel B of FIG. 19 show that body weight and survivalrates between groups of test animals were comparable, thus indicatingthat KD025 has no effect on body weight and survival rates. Survivaldata were analyzed by life-table methods, and actuarial survival ratesare shown. Group comparisons were made by log-rank test.

cGVHD is characterized by fibrosis and increased collagen depositionaround bronchioles and blood vessels in lungs. In addition, elevatedantibody deposition is also observed in lungs. Organs from mice treatedwith donor bone marrow (BM) cells, with or without allogeneicsplenocytes (“vehicle”), and with or without 150 mg/kg of KD025 wereharvested, embedded in optimal cutting temperature compound, snap-frozenin liquid nitrogen, and stored at −80° C. Lungs were inflated byinfusion of 1 mL of optimal cutting temperature compound:PBS (3:1)intratracheally prior to harvest. Six-micrometer cryosections were fixedfor 5 minutes in acetone and stained with mMasson's trichrome stainingkit (Sigma-Aldrich) for detection of collagen deposition. Collagendeposition was quantified on trichrome-stained sections as a ratio ofarea of blue staining to area of total staining by use of the AdobemPhotoshop CS3 analysis tool (Adobe Systems). For measuring Igdeposition, acetone-fixed 6-m lung cryosections were blocked with horseserum and streptavidin-biotin blocking kit (Vector) and stained withfluorescein isothiocyanate (FITC)-labeled anti-mouse-Ig (BD Pharmingen).Antibody deposition was quantified by area of Ig staining per 100-μmsection.

FIG. 20 shows collagen deposition and antibody deposition in lungs oftest mice following transplantation with donor bone marrow (BM) cells,with or without induction by allogeneic splenocytes (“vehicle”), andwith or without KD025-mediated ROCK2 inhibition. Tissue samples fromKD025 treated mice were essentially indistinguishable from samples fromuninduced mice that did not receive allogeneic splenocytes.

Example 215 Reduction of Number of Germinal Centers and Frequency of TFollicular Helper (Tfh) Cells in Spleen of Murine Model of Chronic GVHDby Selective ROCK2 Inhibitor

Germinal centers (GCs) in the spleen are sites of interaction betweenproliferating antigen-specific B cells and T follicular helper (Tfh)cells, where mature B cells rapidly proliferate, differentiate, andundergo somatic hypermutation and selection that results in theproduction of class-switched antibody. Tfh cells are critical for theformation and function of B cell responses. Tfh cells are commonlyidentified by high surface expression of the chemokine receptor CXCR5,and the inhibitory receptor programmed cell death-1 (PD-1). CXCR5expression allows Tfh cells to migrate from the T cell zone to the Bcell follicle where they localize to the GC, and mediate B cell help viacell-cell contact and secretion of the cytokines, such IL-21 and IL-4.Robust GC reactions are present at the time of cGVHD disease initiationand blockade of GC formation has been observed to suppress cGVHD.

Mice were administered 150 mg/kg of KD025 and otherwise treated asdescribed above. Fixed 6-μm spleen cryosections of mice were stainedwith rhodamine-conjugated peanut agglutinin (PNA; Vector Laboratories),and the size of the PNA-positive sections was measured and defined asgerminal centers. For flow cytometric analysis of Tfh cells and GC Bcells, single-cell suspensions from spleens were obtained and labeledwith anti-CD4, anti-CXCR5, and anti-PD1 antibodies (eBioscience). Cellswere analyzed on a BD LSRFortessa cell analyzer.

Spleens of mice transplanted with BM cells and allogeneic splenocytesdemonstrated increased numbers of GCs, as well as increased numbers ofCD4⁺PD1^(Hi)CXCR5 Tfh cells. Both effects were reversed byadministration of KD025 (FIG. 21).

Example 216 KD025 Concurrently Regulates STAT3/STAT5 Phosphorylation andDown-Regulates Protein Levels of RORγt, IRF4 and Bcl6 in a Murine Modelof cGVHD

Mice were treated as described above, except that twelve animals pergroup were tested. Blood was collected on day 56 before or after thelast dose of KD025 by cardiac puncture, and plasma was prepared andstored at −80° C. (n=3/group). Spleens were also collected for analysison day 56. Pulmonary function tests were performed on day 98 (42 dayspost KD025 treatment). Pulmonary function tests and analysis of bodyweight were performed as described above.

Single cell suspensions were prepared from the isolated spleens. Cellswere lysed in RIPA buffer and analyzed for protein content. Samplebuffer was then added, and, after boiling, the samples, containing equalamounts of proteins, were separated by SDS-PAGE gel electrophoresis andtransferred to a nitrocellulose membrane. Membranes were blocked andprobed with specific antibodies overnight. Actin was used as a loadingcontrol. Anti-pSTAT3 pSTAT5 and Bcl6 antibodies were purchased from CellSignaling Technologies (Beverly, Mass.). Anti-IRF4 (sc-48338) andanti-RORγt (14-6988) were obtained from Santa Cruz Biotechnology Inc.and eBioscience respectively. Immunoreactive protein bands werevisualized using an HRP-conjugated secondary antibodies and an enhancedECL system.

Treatment of mice with the selective ROCK2 inhibitor KD025 from days28-56 after transplantation lead to a significant reduction in STAT3phosphorylation and concurrent increase in STAT5 phosphorylation inspleens (FIG. 22, Panel A). In addition, targeted ROCK2 inhibitionresulted in a dose-dependent decrease in protein levels of transcriptionfactors such as RORγt, IRF4 and Bcl6 in spleenocytes (FIG. 22, Panel B).Body weight curves were comparable between groups (FIG. 23). PlasmaKD025 concentrations values were measurable and generally dose dependent(day 56 1-hr pre-dose), and increased with increasing doses and weregenerally at levels consistent with previous experiments (day 56, 1-hrpost-dose) (FIG. 24).

The pulmonary function tests performed on day 98 revealed that KD025treatment leads to long-lasting improvement of respiratory function anddose-dependent decrease in lung resistance was observed 42 days afterthe last administration of KD025 (FIG. 25).

Example 217 Therapeutic Administration of KD025 Blocks Progression ofSclerodermatous cGVHD in Mice

To assess the efficacy of KD025 as a therapeutic intervention for cGVHDa well-characterized B10.D2 (H-2^(d)) into BALB/c (H-2d) MHC-matched,minor histocompatibility Ag-mismatched model was used that recapitulatesthe pathopsysiology of cGVHD, especially the sclerodermatous form of thedisease. In this model, BALB/c (H-2^(d)) mice received 775 cGy of totalbody radiation on day 0 and were transplanted with bone marrow andT-cells from B10.D2 (H-2^(d)) mice. KD025 was administeredintraperitoneally once daily from days 19-47. Compared to vehicletreated mice, KD025 treatment blocked the development of chronicsclerodermatous cGVHD and significantly decreased the GVHD score in thismouse model. The GVHD score in this model combined changes in weightlost, posture, activity, fur texture, and skin integrity (FIG. 26).Sclerodermatous cGVHD mice were further analyzed as depicted in FIGS.27-35.

We claim:
 1. A method of treating sclerodermatous chronic graft versus host disease in a subject in need thereof, which comprises administering to the subject an effective amount of a compound of Formula:

or pharmaceutically acceptable salt thereof.
 2. The method of claim 1, wherein the compound is administered to the subject with a second agent. 